LOCUS       V030027                 9271 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030027
VERSION     V030027
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..3297
                     /label="Plaur(NM_011113)"
                     /note="Plaur(NM_011113)"
                     /gene="Plaur"
     CDS             3304..3369
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3413..3624
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3637..3905
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3934..3943
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3964..4626
                     /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             4696..4749
                     /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             4750..5349
                     /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    5350..5938
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6012..6245
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6317..6438
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6478..6613
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6646..6662)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6670..6686
                     /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(6694..6724)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6739..6760
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7048..7636)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7807..8667)
                     /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(8668..8772)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9246..9262
                     /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 gccaccatgg gactcccaag gcggctgctg
     2341 ctgctgctgt tgctggcgac tacctgtgtc ccagcctccc agggcctgca gtgcatgcag
     2401 tgtgagagta accagagctg cctggtagag gagtgtgctc tgggccagga cctctgcagg
     2461 actaccgtgc ttcgggaatg gcaagatgat agagagctgg aggtggtgac aagaggctgt
     2521 gcccacagcg aaaagaccaa caggaccatg agttaccgca tgggctccat gatcatcagc
     2581 ctgacagaga ccgtgtgcgc cacaaacctc tgcaacaggc ccagacccgg agcccgaggc
     2641 cgtgctttcc cccagggccg ttacctcgag tgtgcgtcct gcacctcttt ggaccagagc
     2701 tgtgagaggg gccgggagca aagcctgcaa tgccgctatc ctacagagca ctgtattgaa
     2761 gtggtgaccc tccagagcac agaaaggagc ttgaaggatg aggactacac ccgaggctgt
     2821 ggcagtcttc ccggatgccc aggcacagca ggtttccata gcaaccagac ctttcacttc
     2881 ctgaagtgtt gcaactacac ccactgcaat ggtggcccag ttctggatct tcagagcttt
     2941 ccaccgaatg gcttccagtg ttacagctgt gaggggaaca atacccttgg gtgttcctcc
     3001 gaagaggcgt ctctcatcaa ctgccgggga ccaatgaatc agtgcctggt ggctacaggc
     3061 ttagatgtgc tgggaaaccg gagttatacc gtaagaggct gcgccacggc ttcctggtgc
     3121 caaggctccc acgtggcaga ctccttcccg acccacctca acgtctctgt ctcctgctgc
     3181 cacggcagcg gctgtaacag ccccacaggg ggcgccccca ggccaggccc tgctcaactc
     3241 agcctcattg cctctctgct cctgaccctc ggactatggg gcgtcctcct ctggaccgga
     3301 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3361 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3421 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     3481 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3541 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3601 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     3661 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     3721 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     3781 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     3841 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     3901 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc
     3961 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag
     4021 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc
     4081 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc
     4141 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac
     4201 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc
     4261 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc
     4321 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag
     4381 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg
     4441 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc
     4501 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg
     4561 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc
     4621 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc
     4681 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     4741 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4801 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4861 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4921 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4981 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5041 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5101 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5161 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5221 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5281 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5341 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5401 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5461 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5521 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5581 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5641 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5701 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     5761 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5821 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5881 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     5941 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6001 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg
     6061 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6121 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6181 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6241 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     6301 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6361 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6421 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     6481 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6541 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6601 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc
     6661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8821 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8881 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     8941 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9001 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9061 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9241 acgttgtaaa acgacggcca gtgccaagct g
//
