LOCUS       V033689                 8711 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033689
VERSION     V033689
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..2805
                     /label="HINT2(NM_032593)"
                     /note="HINT2(NM_032593)"
                     /gene="HINT2"
     promoter        2870..3081
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3094..3362
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3391..3400
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3421..4083
                     /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             4153..4206
                     /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             4207..4806
                     /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    4807..5394
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5468..5701
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5773..5894
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5913..6048
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6086..6102)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6110..6126
                     /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(6134..6164)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6179..6200
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6488..7076)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7247..8107)
                     /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(8108..8212)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8686..8702
                     /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 cggcagccgt ggtgctggct
     2341 gctgggttgc gcgcggcgcg cagagccgtg gcggccacgg gggtgcgcgg ggggcaggtc
     2401 cgaggagctg caggtgtgac tgatgggaat gaagtggcca aggcccagca ggcaactcct
     2461 gggggagcag ccccaaccat cttctcccgg atcctggaca agagcctccc agctgacatt
     2521 ctctatgagg accagcagtg tcttgtgttc cgtgatgtgg cccctcaggc tcctgtgcac
     2581 ttcctggtca ttcctaagaa gcccattcct cggattagcc aggctgaaga agaagaccag
     2641 cagcttctag gacacctact ccttgtggcc aagcagacag caaaggctga gggcctggga
     2701 gatggatacc gacttgtgat caacgatggg aagctgggtg cacaatctgt gtatcatctg
     2761 cacattcatg tacttggggg ccggcagctc cagtggcctc caggttgagg atccctgcga
     2821 tatccagcgc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     2881 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     2941 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     3001 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     3061 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     3121 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     3181 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     3241 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     3301 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     3361 tccaagctgt gaccggcgcc tacgctagac gccaccatgg agagcgacga gagcggcctg
     3421 cccgccatgg agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg
     3481 gtgggcggcg gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc
     3541 aaaggcgccc tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac
     3601 cacttcggca cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc
     3661 ggctacacca acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc
     3721 agctaccgct acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc
     3781 cccgaggaca gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac
     3841 ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc
     3901 gacggcggct actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac
     3961 cccagcatcc tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac
     4021 agcaacaccg agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc
     4081 gccagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc
     4141 accggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     4201 ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4261 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4321 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4381 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4441 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4501 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     4561 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4621 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     4681 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4741 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4801 gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4861 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4921 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4981 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5041 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5101 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5161 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     5221 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     5281 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5341 cgtctccgcc ttcgccctca gacgagtcgg atctcccttt ggccgcctcc ccgcctggta
     5401 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     5461 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     5521 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5581 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5641 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5701 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     5761 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5881 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     5941 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6001 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6061 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6121 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6181 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6241 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6301 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6901 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7141 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7321 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7561 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7861 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7981 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8041 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8101 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8161 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8221 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8281 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8341 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8401 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     8461 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     8521 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     8581 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     8641 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     8701 gtgccaagct g
//
