LOCUS       V043909                 9511 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043909
VERSION     V043909
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..3537
                     /label="METTL8(NM_024770)"
                     /note="METTL8(NM_024770)"
                     /gene="METTL8"
     CDS             3544..3609
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3653..3864
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3877..4145
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4174..4183
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4204..4866
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4936..4989
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4990..5589
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5590..6178
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6252..6485
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6557..6678
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6718..6853
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6886..6902)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6910..6926
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6934..6964)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6979..7000
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7288..7876)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8047..8907)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8908..9012)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9486..9502
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatga atatgatttg gagaaattcc
     2341 atttcttgtc taaggctagg aaaggtgcca cacagatacc aaagtggtta ccacccagtg
     2401 gcccctctgg gatcaaggat tttaactgac ccagccaaag tttttgaaca caacatgtgg
     2461 gatcacatgc agtggtctaa ggaagaagaa gcagcagcca gaaaaaaagt aaaagaaaac
     2521 tcagctgtgc gagtccttct ggaagagcaa gttaagtatg agagagaagc tagtaaatac
     2581 tgggacacat tttacaagat tcataagaat aagtttttca aggatcgtaa ttggctgttg
     2641 agggaatttc ctgaaattct tccagttgat caaaaacctg aagagaaggc gagagaatca
     2701 tcatgggatc atgtaaaaac tagtgctaca aatcgtttct caagaatgca ctgtcctact
     2761 gtgcctgatg aaaaaaatca ttatgagaaa agttctggtt cttcagaagg tcaaagcaaa
     2821 acagaatctg atttttccaa cctagactct gaaaaacaca aaaaaggacc tatggagact
     2881 ggattgtttc ctggtagcaa tgccactttc aggatactag aggttggttg tggagctgga
     2941 aatagtgtgt ttccaatttt gaacactttg gagaactctc cggagtcctt tctgtattgt
     3001 tgtgattttg cttctggagc tgtggagctc gtaaagtcac actcgtccta cagagcaacc
     3061 cagtgttttg cctttgttca tgatgtatgt gatgatggct taccttaccc ttttccagat
     3121 gggatcctgg atgtcattct ccttgtcttt gtgctctctt ctattcatcc tgacaggatg
     3181 caaggtgttg taaaccgact gtccaagtta ctgaaacctg ggggaatgct gttatttcga
     3241 gactatggaa gatatgataa gactcagctt cgttttaaaa agggacattg tttatctgaa
     3301 aatttttatg ttcgaggaga tggtaccaga gcatatttct ttacaaaagg ggaagtccac
     3361 agtatgttct gcaaagccag tttagatgaa aagcaaaatc tggttgatcg ccgcttacaa
     3421 gttaatagga aaaaacaagt gaaaatgcac cgagtgtgga ttcaaggcaa attccagaaa
     3481 ccattgcacc agactcagaa tagctccaat atggtatcta cactcctttc acaagacgga
     3541 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3601 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3661 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     3721 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3781 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3841 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     3901 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     3961 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     4021 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4081 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     4141 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc
     4201 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag
     4261 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc
     4321 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc
     4381 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac
     4441 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc
     4501 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc
     4561 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag
     4621 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg
     4681 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc
     4741 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg
     4801 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc
     4861 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc
     4921 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     4981 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5041 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5101 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5161 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5221 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5281 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5341 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5401 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5461 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5521 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5581 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5641 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5701 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5761 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5821 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5881 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5941 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     6001 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     6061 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     6121 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     6181 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6241 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg
     6301 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6361 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6421 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6481 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     6541 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6601 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6661 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     6721 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6781 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6841 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc
     6901 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6961 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7081 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8341 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8401 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8461 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8521 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8581 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8641 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8701 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8761 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8821 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8881 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8941 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9001 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9061 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9121 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9181 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9241 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9301 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9481 acgttgtaaa acgacggcca gtgccaagct g
//
