LOCUS       V037988                 9555 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037988
VERSION     V037988
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             2302..3660
                     /label="DNAH14(NM_001145154)"
                     /note="DNAH14(NM_001145154)"
                     /gene="DNAH14"
     promoter        3714..3925
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3938..4206
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4235..4244
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4265..4927
                     /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             4997..5050
                     /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             5051..5650
                     /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    5651..6238
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6312..6545
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6617..6738
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6757..6892
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6930..6946)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6954..6970
                     /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(6978..7008)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7023..7044
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7332..7920)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8091..8951)
                     /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(8952..9056)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9530..9546
                     /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 tagaagatta
     2281 atactttcag aattcgccac catggagacg tttataccca ttgatttgac aactgaaaat
     2341 caagagatgg acaaggagga aaccaagaca aaaccaagac ttttaagata tgaagagaaa
     2401 aaatatgaag atgtgaaacc attagagact caaccagctg aaatagcaga aaaggaaaca
     2461 ttggaatata aaacagttag aacattctct gaatctttga agtcagagaa aacagaagat
     2521 taccttagag aaagtataat tcaacaacat atggtttctc cagagccagc ttcccttaag
     2581 gagaaaggga agtcaaggag aaaaaaggat caaactcatg cttgtccaaa tgttaggaaa
     2641 gccaggcctg tgtcctatga tagaacagaa ccaaaagatg atgatgtgat aagaaatatt
     2701 attaggctac gagaaaagct tggttggcaa actatattac cgcagcacag tttgaaatac
     2761 ggaagctcca aaattgcaat tcagaagatt actttaaaga aacctttgga agatgatgga
     2821 gaatttgttt attgccttcc tcggaaaagt cctaaatccc tttacaatcc atatgatctt
     2881 caggtagtat cggctcatac tgctaaacat tgcaaagaat tttgggttat tactgcttca
     2941 tttatctcaa aggttattaa tatagttggt agtgtaaagg aagtagaact catacctact
     3001 ttggaatggc tatcagaaag aagacattac tatttattac ggcaattcaa gatattttct
     3061 gatttccgaa tgaataaagc atttgttacc tggaaattga atgttaaaag aattaagaca
     3121 gagaagagca ggtcattttt gtaccaccat ctttttttgg ctgatgactt gtttcaaacc
     3181 tgtttggttt atataagagg actttgtgaa gatgcaatta atctcaaaaa ttataatgac
     3241 catgaaaata atctatctgc catatgcctt gtaaagctgg atagttctcg aacatattct
     3301 ctagatgaat tttgtgaaga gcagttacag caagctaccc aggcattgaa acaacttgag
     3361 gacatcagga ataaagcaat ttcagagatg aaaagtactt ttctaaaggt tgcagaaaag
     3421 aatgaaatca aagagtattt tgagtcaaaa ctctctgaag atgacacaac acatttcaag
     3481 ctgcctaaat atagacgttt attagaaaca tttttcaagt ttgtaatgct ggttgactac
     3541 atatttcagg aactcattcg tcaacttatg aacactgcag tcacactact tttggaatta
     3601 tttaatggtt ctgctggaat gccattttca gtggaaaaaa agaatgaaaa tcttatcagg
     3661 taactcgagt aagaacaact gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     3721 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     3781 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     3841 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     3901 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     3961 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     4021 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     4081 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     4141 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     4201 cagatccaag ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg
     4261 cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga
     4321 gctggtgggc ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag
     4381 caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt
     4441 ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa
     4501 cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag
     4561 cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg
     4621 cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga
     4681 gcacctgcac cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct
     4741 gcgcgacggc ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat
     4801 ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct
     4861 gcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc
     4921 cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg
     4981 ctccaccgga tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     5041 tcccggccct atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     5101 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     5161 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     5221 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     5281 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     5341 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     5401 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     5461 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     5521 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     5581 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     5641 cggtgcctga aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5701 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5761 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5821 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5881 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5941 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     6001 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     6061 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6121 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6181 tccgcgtctc cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct
     6241 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6301 aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg
     6361 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6421 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6481 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6541 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     6601 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6661 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6721 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt
     6781 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6841 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt
     6901 tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6961 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7021 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7081 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7141 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7201 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7501 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7741 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7921 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7981 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8041 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8101 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8161 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8221 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8281 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8341 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8401 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8461 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8521 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8581 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8641 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8701 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8761 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8821 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8881 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8941 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9001 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9061 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     9121 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     9181 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     9241 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     9301 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     9361 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     9421 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     9481 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     9541 gccagtgcca agctg
//
