LOCUS       V023760                 9465 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023760
VERSION     V023760
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             2303..3580
                     /label="INHBA(NM_002192)"
                     /note="INHBA(NM_002192)"
                     /gene="INHBA"
     promoter        3624..3835
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3848..4116
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4145..4154
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4175..4837
                     /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             4907..4960
                     /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             4961..5560
                     /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    5561..6148
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6222..6455
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6527..6648
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6667..6802
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6840..6856)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6864..6880
                     /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(6888..6918)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6933..6954
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7242..7830)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8001..8861)
                     /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(8862..8966)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9440..9456
                     /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 gaattcgcca ccatgccctt gctttggctg agaggatttc tgttggcaag
     2341 ttgctggatt atagtgagga gttcccccac cccaggatcc gaggggcaca gcgcggcccc
     2401 cgactgtccg tcctgtgcgc tggccgccct cccaaaggat gtacccaact ctcagccaga
     2461 gatggtggag gccgtcaaga agcacatttt aaacatgctg cacttgaaga agagacccga
     2521 tgtcacccag ccggtaccca aggcggcgct tctgaacgcg atcagaaagc ttcatgtggg
     2581 caaagtcggg gagaacgggt atgtggagat agaggatgac attggaagga gggcagaaat
     2641 gaatgaactt atggagcaga cctcggagat catcacgttt gccgagtcag gaacagccag
     2701 gaagacgctg cacttcgaga tttccaagga aggcagtgac ctgtcagtgg tggagcgtgc
     2761 agaagtctgg ctcttcctaa aagtccccaa ggccaacagg accaggacca aagtcaccat
     2821 ccgcctcttc cagcagcaga agcacccgca gggcagcttg gacacagggg aagaggccga
     2881 ggaagtgggc ttaaaggggg agaggagtga actgttgctc tctgaaaaag tagtagacgc
     2941 tcggaagagc acctggcatg tcttccctgt ctccagcagc atccagcggt tgctggacca
     3001 gggcaagagc tccctggacg ttcggattgc ctgtgagcag tgccaggaga gtggcgccag
     3061 cttggttctc ctgggcaaga agaagaagaa agaagaggag ggggaaggga aaaagaaggg
     3121 cggaggtgaa ggtggggcag gagcagatga ggaaaaggag cagtcgcaca gacctttcct
     3181 catgctgcag gcccggcagt ctgaagacca ccctcatcgc cggcgtcggc ggggcttgga
     3241 gtgtgatggc aaggtcaaca tctgctgtaa gaaacagttc tttgtcagtt tcaaggacat
     3301 cggctggaat gactggatca ttgctccctc tggctatcat gccaactact gcgagggtga
     3361 gtgcccgagc catatagcag gcacgtccgg gtcctcactg tccttccact caacagtcat
     3421 caaccactac cgcatgcggg gccatagccc ctttgccaac ctcaaatcgt gctgtgtgcc
     3481 caccaagctg agacccatgt ccatgttgta ctatgatgat ggtcaaaaca tcatcaaaaa
     3541 ggacattcag aacatgatcg tggaggagtg tgggtgctca tagcggccgc gaaggatctg
     3601 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     3661 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     3721 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     3781 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     3841 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     3901 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     3961 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     4021 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     4081 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct
     4141 agacgccacc atggagagcg acgagagcgg cctgcccgcc atggagatcg agtgccgcat
     4201 caccggcacc ctgaacggcg tggagttcga gctggtgggc ggcggagagg gcacccccaa
     4261 gcagggccgc atgaccaaca agatgaagag caccaaaggc gccctgacct tcagccccta
     4321 cctgctgagc cacgtgatgg gctacggctt ctaccacttc ggcacctacc ccagcggcta
     4381 cgagaacccc ttcctgcacg ccatcaacaa cggcggctac accaacaccc gcatcgagaa
     4441 gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg ccggccgcgt
     4501 gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga tcttcaccga
     4561 caagatcatc cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg ataacgtgct
     4621 ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc ggctactaca gcttcgtggt
     4681 ggacagccac atgcacttca agagcgccat ccaccccagc atcctgcaga acgggggccc
     4741 catgttcgcc ttccgccgcg tggaggagct gcacagcaac accgagctgg gcatcgtgga
     4801 gtaccagcac gccttcaaga cccccatcgc cttcgccaga tcccgcgctc agtcgtccaa
     4861 ttctgccgtg gacggcaccg ccggacccgg ctccaccgga tctcgcgagg gcagaggaag
     4921 tcttctaaca tgcggtgacg tggaggagaa tcccggccct atgaccgagt acaagcccac
     4981 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     5041 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     5101 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5161 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5221 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5281 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5341 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5401 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5461 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5521 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga aatcaacctc tggattacaa
     5581 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5641 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5701 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5761 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5821 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5881 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5941 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     6001 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6061 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc cgccttcgcc ctcagacgag
     6121 tcggatctcc ctttggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc
     6181 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     6241 ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     6301 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6361 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6421 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     6481 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     6541 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6601 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     6661 ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6721 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6781 tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg
     6841 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6901 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6961 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7021 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7081 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7141 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7201 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7261 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7321 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7381 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7441 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7501 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7561 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7621 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7681 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7741 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7801 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7861 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7921 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7981 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8041 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8101 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8161 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8221 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8281 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8341 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8401 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8461 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8521 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8581 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8641 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8701 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8761 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8821 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     9001 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     9061 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     9121 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     9181 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     9241 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     9301 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     9361 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     9421 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
