LOCUS       V044162                 9244 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044162
VERSION     V044162
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..3270
                     /label="APEX1(NM_001641)"
                     /note="APEX1(NM_001641)"
                     /gene="APEX1"
     CDS             3277..3342
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3386..3597
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3610..3878
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3907..3916
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3937..4599
                     /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             4669..4722
                     /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             4723..5322
                     /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    5323..5911
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5985..6218
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6290..6411
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6451..6586
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6619..6635)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6643..6659
                     /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(6667..6697)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6712..6733
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7021..7609)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7780..8640)
                     /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(8641..8745)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9219..9235
                     /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 gccaccatgc cgaagcgtgg gaaaaaggga
     2341 gcggtggcgg aagacgggga tgagctcagg acagagccag aggccaagaa gagtaagacg
     2401 gccgcaaaga aaaatgacaa agaggcagca ggagagggcc cagccctgta tgaggacccc
     2461 ccagatcaga aaacctcacc cagtggcaaa cctgccacac tcaagatctg ctcttggaat
     2521 gtggatgggc ttcgagcctg gattaagaag aaaggattag attgggtaaa ggaagaagcc
     2581 ccagatatac tgtgccttca agagaccaaa tgttcagaga acaaactacc agctgaactt
     2641 caggagctgc ctggactctc tcatcaatac tggtcagctc cttcggacaa ggaagggtac
     2701 agtggcgtgg gcctgctttc ccgccagtgc ccactcaaag tttcttacgg cataggcgat
     2761 gaggagcatg atcaggaagg ccgggtgatt gtggctgaat ttgactcgtt tgtgctggta
     2821 acagcatatg tacctaatgc aggccgaggt ctggtacgac tggagtaccg gcagcgctgg
     2881 gatgaagcct ttcgcaagtt cctgaagggc ctggcttccc gaaagcccct tgtgctgtgt
     2941 ggagacctca atgtggcaca tgaagaaatt gaccttcgca accccaaggg gaacaaaaag
     3001 aatgctggct tcacgccaca agagcgccaa ggcttcgggg aattactgca ggctgtgcca
     3061 ctggctgaca gctttaggca cctctacccc aacacaccct atgcctacac cttttggact
     3121 tatatgatga atgctcgatc caagaatgtt ggttggcgcc ttgattactt tttgttgtcc
     3181 cactctctgt tacctgcatt gtgtgacagc aagatccgtt ccaaggccct cggcagtgat
     3241 cactgtccta tcaccctata cctagcactg ggatccgact acaaagacca tgacggtgat
     3301 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc
     3361 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     3421 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     3481 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3541 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     3601 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     3661 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     3721 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     3781 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     3841 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     3901 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc
     3961 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc
     4021 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc
     4081 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc
     4141 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag
     4201 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc
     4261 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc
     4321 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg
     4381 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg
     4441 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc
     4501 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg
     4561 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc
     4621 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga
     4681 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc
     4741 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     4801 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     4861 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4921 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4981 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     5041 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     5101 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5161 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5221 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5281 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac
     5341 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     5401 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     5461 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     5521 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     5581 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     5641 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     5701 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     5761 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     5821 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     5881 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa
     5941 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     6001 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     6061 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6121 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6181 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     6241 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     6301 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6361 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6421 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6481 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6541 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga
     6601 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6661 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6721 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6781 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7441 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8761 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8821 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     8881 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     8941 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     9001 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     9061 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     9121 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     9181 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     9241 gctg
//
