LOCUS       V027253                 9657 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027253
VERSION     V027253
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2301..4511
                     /label="PROX1(NM_002763)"
                     /note="PROX1(NM_002763)"
                     /gene="PROX1"
     CDS             4518..4583
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4627..4838
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4851..5119
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5146..5745
                     /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    5752..6340
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6414..6647
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6719..6840
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6859..6994
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7032..7048)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7056..7072
                     /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(7080..7110)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7125..7146
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7434..8022)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8193..9053)
                     /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(9054..9158)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9632..9648
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgcctgacc atgacagcac agccctctta agccggcaaa
     2341 ccaagaggag aagagttgac attggagtga aaaggacggt agggacagca tctgcatttt
     2401 ttgctaaggc aagagcaacg ttttttagtg ccatgaatcc ccaaggttct gagcaggatg
     2461 ttgagtattc agtggtgcag catgcagatg gggaaaagtc aaatgtactc cgcaagctgc
     2521 tgaagagggc gaactcgtat gaagatgcca tgatgccttt tccaggagca accataattt
     2581 cccagctgtt gaaaaataac atgaacaaaa atggtggcac ggagcccagt ttccaagcca
     2641 gcggtctctc tagtacaggc tccgaagtac atcaggagga tatatgcagc aactcttcaa
     2701 gagacagccc cccagagtgt ctttcccctt ttggcaggcc tactatgagc cagtttgata
     2761 tggatcgctt atgtgatgag cacctgagag caaagcgcgc ccgggttgag aatataattc
     2821 ggggtatgag ccattccccc agtgtggcat taaggggcaa tgaaaatgaa agagagatgg
     2881 ccccgcagtc tgtgagtccc cgagaaagtt acagagaaaa caaacgcaag caaaagcttc
     2941 cccagcagca gcaacagagt ttccagcagc tggtttcagc ccgaaaagaa cagaagcgag
     3001 aggagcgccg acagctgaaa cagcagctgg aggacatgca gaaacagctg cgccagctgc
     3061 aggaaaagtt ctaccaaatc tatgacagca ctgattcgga aaatgatgaa gatggtaacc
     3121 tgtctgaaga cagcatgcgc tcggagatcc tggatgccag ggcccaggac tctgtcggaa
     3181 ggtcagataa tgagatgtgc gagctagacc caggacagtt tattgaccga gctcgagccc
     3241 tgatcagaga gcaggaaatg gctgaaaaca agccgaagcg agaaggcaac aacaaagaaa
     3301 gagaccatgg gccaaactcc ttacaaccgg aaggcaaaca tttggctgag accttgaaac
     3361 aggaactgaa cactgccatg tcgcaagttg tggacactgt ggtcaaagtc ttttcggcca
     3421 agccctcccg ccaggttcct caggtcttcc cacctctcca gatcccccag gccagatttg
     3481 cagtcaatgg ggaaaaccac aatttccaca ccgccaacca gcgcctgcag tgctttggcg
     3541 acgtcatcat tccgaacccc ctggacacct ttggcaatgt gcagatggcc agttccactg
     3601 accagacaga agcactgccc ctggttgtcc gcaaaaactc ctctgaccag tctgcctccg
     3661 gccctgccgc tggcggccac caccagcccc tgcaccagtc gcctctctct gccaccacgg
     3721 gcttcaccac gtccaccttc cgccacccct tcccccttcc cttgatggcc tatccatttc
     3781 agagcccatt aggtgctccc tccggctcct tctctggaaa agacagagcc tctcctgaat
     3841 ccttagactt aactagggat accacgagtc tgaggaccaa gatgtcatct caccacctga
     3901 gccaccaccc ttgttcacca gcacacccgc ccagcaccgc cgaagggctc tccttgtcgc
     3961 tcataaagtc cgagtgcggc gatcttcaag atatgtctga aatatcacct tattcgggaa
     4021 gtgcaatgca ggaaggattg tcacccaatc acttgaaaaa agcaaagctc atgttttttt
     4081 atacccgtta tcccagctcc aatatgctga agacctactt ctccgacgta aagttcaaca
     4141 gatgcattac ctctcagctc atcaagtggt ttagcaattt ccgtgagttt tactacattc
     4201 agatggagaa gtacgcacgt caagccatca acgatggggt caccagtact gaagagctgt
     4261 ctataaccag agactgtgag ctgtacaggg ctctgaacat gcactacaat aaagcaaatg
     4321 actttgaggt tccagagaga ttcctggaag ttgctcagat cacattacgg gagtttttca
     4381 atgccattat cgcaggcaaa gatgttgatc cttcctggaa gaaggccata tacaaggtca
     4441 tctgcaagct ggatagtgaa gtccctgaga ttttcaaatc cccgaactgc ctacaagagc
     4501 tgcttcatga gggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     4561 actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc
     4621 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     4681 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     4741 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     4801 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     4861 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     4921 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     4981 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     5041 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     5101 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag
     5161 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     5221 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     5281 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     5341 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5401 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     5461 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     5521 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5581 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     5641 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     5701 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct
     5761 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5821 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5881 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5941 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     6001 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     6061 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     6121 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     6181 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6241 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     6301 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat
     6361 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6421 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt
     6481 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6541 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6601 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     6661 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     6721 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6781 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6841 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     6901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6961 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat
     7021 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7081 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7141 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7201 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7261 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7321 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7381 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7441 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7501 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7561 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7621 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7681 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7741 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7801 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7861 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7921 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7981 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8041 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8101 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8161 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8761 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8821 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8881 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8941 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     9001 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     9061 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9121 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9181 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     9241 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     9301 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     9361 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     9421 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     9481 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     9541 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     9601 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
