LOCUS       V032699                10208 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032699
VERSION     V032699
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             2287..2352
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2377..5133
                     /label="TJP3(NM_001267560)"
                     /note="TJP3(NM_001267560)"
                     /gene="TJP3"
     promoter        5178..5389
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5402..5670
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5697..6296
                     /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    6303..6891
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6965..7198
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7270..7391
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7410..7545
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7583..7599)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7607..7623
                     /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(7631..7661)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7676..7697
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7985..8573)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8744..9604)
                     /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(9605..9709)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10183..10199
                     /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 agaatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     2341 gacgatgaca agtctagaga attcggatcc gccaccatgg aggagctgac catctgggaa
     2401 cagcacacgg ccacactgtc caaggacccc cgccggggct ttggcattgc gatctctgga
     2461 ggccgagacc ggcccggtgg atccatggtt gtatctgacg tggtacctgg agggccggcg
     2521 gagggcaggc tacagacagg cgaccacatc gtcatggtga acggggtttc catggagaat
     2581 gccacctccg cgtttgccat tcagatactc aagacctgca ccaagatggc caacatcaca
     2641 gtgaaacgtc cccggaggat ccacctgccc gccaccaaag ccagcccctc cagcccaggg
     2701 cgccaggact cggatgaaga cgatgggccc cagcgggtgg aggaggtgga ccagggccgg
     2761 ggctatgacg gcgactcatc cagtggctcc ggccgctcct gggacgagcg ctcccgccgg
     2821 ccgaggcctg gtcgccgggg ccgggccggc agccatgggc gtaggagccc aggtggtggc
     2881 tctgaggcca acgggctggc cctggtgtcc ggctttaagc ggctgccacg gcaggacgtg
     2941 cagatgaagc ctgtgaagtc agtgctggtg aagaggagag acagcgaaga gtttggcgtc
     3001 aagctgggca gtcagatctt catcaagcac attacagatt cgggcctggc tgcccggcac
     3061 cgtgggctgc aggaaggaga tctcattcta cagatcaacg gggtgtctag ccagaacctg
     3121 tcactgaacg acacccggcg actgattgag aagtcagaag ggaagctaag cctgctggtg
     3181 ctgagagatc gtgggcagtt cctggtgaac attccgcctg ctgtcagtga cagcgacagc
     3241 tcgccattgg aggacatctc ggacctcgcc tcggagctat cgcaggcacc accatcccac
     3301 atcccaccac caccccggca tgctcagcgg agccccgagg ccagccagac cgactctccc
     3361 gtggagagtc cccggcttcg gcgggaaagt tcagtagatt ccagaaccat ctcggaacca
     3421 gatgagcaac ggtcagagtt gcccagggaa agcagctatg acatctacag agtgcccagc
     3481 agtcagagca tggaggatcg tgggtacagc cccgacacgc gtgtggtccg cttcctcaag
     3541 ggcaagagca tcgggctgcg gctggcaggg ggcaatgacg tgggcatctt cgtgtccggg
     3601 gtgcaggcgg gcagcccggc cgacgggcag ggcatccagg agggagatca gattctgcag
     3661 gtgaatgacg tgccattcca gaacctgaca cgggaggagg cagtgcagtt cctgctgggg
     3721 ctgccaccag gcgaggagat ggagctggtg acgcagagga agcaggacat tttctggaaa
     3781 atggtgcagt cccgcgtggg tgactccttc tacatccgca ctcactttga gctggagccc
     3841 agtccaccgt ctggcctggg cttcacccgt ggcgacgtct tccacgtgct ggacacgctg
     3901 caccccggcc ccgggcagag ccacgcacga ggaggccact ggctggcggt gcgcatgggt
     3961 cgtgacctgc gggagcaaga gcggggcatc attcccaacc agagcagggc ggagcagctg
     4021 gccagcctgg aagctgccca gagggccgtg ggagtcgggc ccggctcctc cgcgggctcc
     4081 aatgctcggg ccgagttctg gcggctgcgg ggtcttcgtc gaggagccaa gaagaccact
     4141 cagcggagcc gtgaggacct ctcagctctg acccgacagg gccgctaccc gccctacgaa
     4201 cgagtggtgt tgcgagaagc cagtttcaag cgcccggtag tgatcctggg acccgtggcc
     4261 gacattgcta tgcagaagtt gactgctgag atgcctgacc agtttgaaat cgcagagact
     4321 gtgtccagga ccgacagccc ctccaagatc atcaaactag acaccgtgcg ggtgattgca
     4381 gaaaaagaca agcatgcgct cctggatgtg accccctccg ccatcgagcg cctcaactat
     4441 gtgcagtact accccattgt ggtcttcttc atccccgaga gccggccggc cctcaaggca
     4501 ctgcgccagt ggctggcgcc tgcctcccgc cgcagcaccc gtcgcctcta cgcacaagcc
     4561 cagaagctgc gaaaacacag cagccacctc ttcacagcca ccatccctct gaatggcacg
     4621 agtgacacct ggtaccagga gctcaaggcc atcattcgag agcagcagac gcggcccatc
     4681 tggacggcgg aagatcagct ggatggctcc ttggaggaca acctagacct ccctcaccac
     4741 ggcctggccg acagctccgc tgacctcagc tgcgacagcc gcgttaacag cgactacgag
     4801 acggacggcg agggcggcgc gtacacggat ggcgagggct acacagacgg cgaggggggg
     4861 ccctacacgg atgtggatga tgagcccccg gctccagccc tggcccggtc ctcggagccc
     4921 gtgcaggcag atgagtccca gagcccgagg gatcgtggga gaatctcggc tcatcagggg
     4981 gcccaggtgg acagccgcca cccccaggga cagtggcgac aggacagcat gcgaacctat
     5041 gaacgggaag ccctgaagaa aaagtttatg cgagtacatg atgcggagtc ctccgatgaa
     5101 gacggctatg actggggtcc ggccactgac ctgtgagcgg ccgcgaagga tctgcgatcg
     5161 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     5221 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     5281 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     5341 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     5401 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     5461 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     5521 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     5581 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     5641 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga
     5701 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     5761 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     5821 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     5881 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     5941 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     6001 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     6061 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     6121 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     6181 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     6241 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg
     6301 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     6361 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     6421 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     6481 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     6541 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     6601 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     6661 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     6721 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6781 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6841 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct
     6901 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     6961 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg
     7021 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     7081 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     7141 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     7201 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     7261 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     7321 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     7381 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc
     7441 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     7501 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag
     7561 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7621 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7681 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8401 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     8461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9721 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     9781 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     9841 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     9901 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     9961 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
    10021 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
    10081 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
    10141 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
    10201 ccaagctg
//
