LOCUS       V045564                 9431 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045564
VERSION     V045564
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..3525
                     /label="Krt19(NM_008471)"
                     /note="Krt19(NM_008471)"
                     /gene="Krt19"
     promoter        3590..3801
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3814..4082
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4111..4120
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4141..4803
                     /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             4873..4926
                     /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             4927..5526
                     /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    5527..6114
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6188..6421
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6493..6614
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6633..6768
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6806..6822)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6830..6846
                     /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(6854..6884)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6899..6920
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7208..7796)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7967..8827)
                     /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(8828..8932)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9406..9422
                     /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 cttcctatag ctatcgccag
     2341 acctcagcta tgtcttcctt tgggggtacg ggcgggggtt cagtacgcat tgggtcaggg
     2401 ggtgttttcc gcgcacccag catccacggg ggctcaggtg gccgcggcgt gtccgtgtcc
     2461 tccacccgct tcgtgacctc gtcctccggg agctatggcg gagtccgcgg tggaagtttt
     2521 agtgggaccc tggctgtgtc tgatgggctg ctgtctggca atgagaagat caccatgcaa
     2581 aacctcaatg atcgtctcgc ctcctacttg gacaaggtgc gcgccctaga gcaggccaat
     2641 ggcgagctgg aggtgaagat ccgcgactgg taccagaagc agggacccgg accctcccga
     2701 gattacaacc actactttaa gaccatcgag gacttgcgcg acaagattct tggtgccacc
     2761 attgacaact ccaagattgt cctacagatt gacaatgctc gcctggctgc agatgacttc
     2821 agaaccaagt ttgagacaga acacgccttg cgtctgagcg tggaggccga catcaacggc
     2881 ctgcgccggg tgctggatga gctgactctg gccaggactg acctggagat gcagattgag
     2941 agcctgaagg aggagctggc ctacctgaag aagaaccatg aggaggaaat tactgccctg
     3001 aggagccagg tgggtggcca ggtcagtgtg gaggtggatt ccactcccgg tgtcgaccta
     3061 gccaagatcc tgagtgagat gagaagtcag tatgagatca tggccgagaa gaaccggaag
     3121 gatgctgaag ccacctacct tgctcggatt gaggagctga acacccaggt cgccgtccac
     3181 tctgagcaga tccagataag caagaccgaa gtcacggacc ttcgacggac cctccagggc
     3241 cttgagattg agctgcagtc ccagctcagc atgaaagctg ccctggaagg cacgctggca
     3301 gagacggagg cccgttatgg agtccagctg tcacagatcc agagcgtgat cagcggtttt
     3361 gaagcccagc tgagcgacgt gcgtgccgac atagagcgcc agaaccagga gtataagcag
     3421 ctcatggaca tcaagtccag gctggagcag gagatcgcca cctaccgcag cctgctggag
     3481 ggccaggaag cccactacaa caatctgccc acccccaagg ccatctgagg atccctgcga
     3541 tatccagcgc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     3601 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     3661 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     3721 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     3781 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     3841 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     3901 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     3961 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4021 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4081 tccaagctgt gaccggcgcc tacgctagac gccaccatgg agagcgacga gagcggcctg
     4141 cccgccatgg agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg
     4201 gtgggcggcg gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc
     4261 aaaggcgccc tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac
     4321 cacttcggca cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc
     4381 ggctacacca acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc
     4441 agctaccgct acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc
     4501 cccgaggaca gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac
     4561 ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc
     4621 gacggcggct actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac
     4681 cccagcatcc tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac
     4741 agcaacaccg agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc
     4801 gccagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc
     4861 accggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     4921 ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4981 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     5041 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5101 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5161 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5221 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     5281 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5341 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     5401 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5461 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5521 gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5581 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5641 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5701 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5761 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5821 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5881 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     5941 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     6001 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6061 cgtctccgcc ttcgccctca gacgagtcgg atctcccttt ggccgcctcc ccgcctggta
     6121 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     6181 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     6241 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6301 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6361 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6421 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6481 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6601 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     6661 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6721 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6781 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6841 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6901 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6961 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7021 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7081 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7141 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7201 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7261 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7321 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7381 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7441 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7501 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7561 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7621 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7681 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7741 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7801 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7861 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7921 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7981 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8041 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8101 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8161 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8221 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8281 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8341 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8401 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8461 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8521 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8581 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8641 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8701 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8761 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8821 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8881 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8941 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     9001 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     9061 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     9121 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     9181 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     9241 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     9301 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     9361 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     9421 gtgccaagct g
//
