LOCUS       V032839                 8641 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032839
VERSION     V032839
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             2302..3495
                     /label="KISS1R(NM_032551)"
                     /note="KISS1R(NM_032551)"
                     /gene="KISS1R"
     CDS             3502..3567
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3611..3822
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3835..4103
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4130..4729
                     /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    4736..5324
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5398..5631
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5703..5824
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5843..5978
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6016..6032)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6040..6056
                     /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(6064..6094)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6109..6130
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6418..7006)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7177..8037)
                     /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(8038..8142)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8616..8632
                     /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgcacacc gtggctacgt ccggacccaa cgcgtcctgg
     2341 ggggcaccgg ccaacgcctc cggctgcccg ggctgtggcg ccaacgcctc ggacggccca
     2401 gtcccttcgc cgcgggccgt ggacgcctgg ctcgtgccgc tcttcttcgc ggcgctgatg
     2461 ctgctgggcc tggtggggaa ctcgctggtc atctacgtca tctgccgcca caagccgatg
     2521 cggaccgtga ccaacttcta catcgccaac ctggcggcca cggacgtgac cttcctcctg
     2581 tgctgcgtcc ccttcacggc cctgctgtac ccgctgcccg gctgggtgct gggcgacttc
     2641 atgtgcaagt tcgtcaacta catccagcag gtctcggtgc aggccacgtg tgccactctg
     2701 accgccatga gtgtggaccg ctggtacgtg acggtgttcc cgttgcgcgc cctgcaccgc
     2761 cgcacgcccc gcctggcgct ggctgtcagc ctcagcatct gggtaggctc tgcggcggtg
     2821 tctgcgccgg tgctcgccct gcaccgcctg tcacccgggc cgcgcgccta ctgcagtgag
     2881 gccttcccca gccgcgccct ggagcgcgcc ttcgcactgt acaacctgct ggcgctgtac
     2941 ctgctgccgc tgctcgccac ctgcgcctgc tatgcggcca tgctgcgcca cctgggccgg
     3001 gtcgccgtgc gccccgcgcc cgccgatagc gccctgcagg ggcaggtgct ggcagagcgc
     3061 gcaggcgccg tgcgggccaa ggtctcgcgg ctggtggcgg ccgtggtcct gctcttcgcc
     3121 gcctgctggg gccccatcca gctgttcctg gtgctgcagg cgctgggccc cgcgggctcc
     3181 tggcacccac gcagctacgc cgcctacgcg cttaagacct gggctcactg catgtcctac
     3241 agcaactccg cgctgaaccc gctgctctac gccttcctgg gctcgcactt ccgacaggcc
     3301 ttccgccgcg tctgcccctg cgcgccgcgc cgcccccgcc gcccccgccg gcccggaccc
     3361 tcggaccccg cagccccaca cgcggagctg ctccgcctgg ggtcccaccc ggcccccgcc
     3421 agggcgcaga agccagggag cagtgggctg gccgcgcgcg ggctgtgcgt cctgggggag
     3481 gacaacgccc ctctcggatc cgactacaaa gaccatgacg gtgattataa agatcatgac
     3541 atcgactaca aggatgacga tgacaagtag cggccgcgaa ggatctgcga tcgctccggt
     3601 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     3661 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     3721 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     3781 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     3841 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     3901 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     3961 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     4021 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     4081 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga tgaccgagta
     4141 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     4201 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     4261 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4321 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4381 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4441 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4501 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4561 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4621 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     4681 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca
     4741 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4801 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4861 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4921 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4981 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5041 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5101 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     5161 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5221 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     5281 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     5341 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5401 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     5461 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5521 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5581 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     5641 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     5701 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5761 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5821 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     5881 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5941 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     6001 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6061 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6121 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6181 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6241 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6301 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6421 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6481 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6601 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6841 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6961 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7021 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7081 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7141 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7201 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7261 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7321 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7381 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7441 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7501 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7561 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7621 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7681 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7741 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7801 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7861 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7921 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7981 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8041 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8101 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8161 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8221 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     8281 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     8341 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     8401 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     8461 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     8521 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     8581 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     8641 g
//
