LOCUS       V018688                 9947 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018688
VERSION     V018688
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"
     regulatory      2290..2299
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2299..2322
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2323..4440
                     /label="TCF12(NM_001322151)"
                     /note="TCF12(NM_001322151)"
                     /gene="TCF12"
     promoter        4490..4701
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4714..4982
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5010..6035
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    6042..6630
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6704..6937
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7009..7130
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7149..7284
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7322..7338)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7346..7362
                     /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(7370..7400)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7415..7436
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7724..8312)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8483..9343)
                     /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(9344..9448)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9922..9938
                     /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 ccaccatgga ttacaaggat gacgacgata agatgaatcc ccagcaacaa
     2341 cgcatggccg ctatagggac cgacaaggag ctgagcgacc tactggactt cagtgcgatg
     2401 ttttccccac ctgttaatag tgggaaaact agaccaacta cactgggaag cagtcaattc
     2461 agtggatcag gtattgatga aagaggaggt acaacatctt ggggaacaag tggtcaacca
     2521 agtccttcct atgattcatc tagaggtttt acagacagcc ctcattacag tgatcacttg
     2581 aatgacagtc gattaggagc ccatgaaggc ttgtccccaa cacctttcat gaactcaaat
     2641 ctgatgggaa aaacatcaga gagaggctca ttttccctgt acagcagaga tactggatta
     2701 ccaggctgtc aatctagtct cctgagacaa gatctggggc ttgggagccc agcacagcta
     2761 tcttcttcag gaaaacctgg gacagcatac tattcattct ctgctacaag ttccaggagg
     2821 agaccactcc atgactctgc agcgcttgat cccttgcaag caaaaaaagt cagaaaggtg
     2881 cctcctggtt tgccttcttc tgtatatgca ccatccccaa attcagatga tttcaaccgt
     2941 gaatctccta gttatccatc tcctaagcca ccaaccagta tgttcgctag cactttcttt
     3001 atgcaagatg ggacccacaa ttcttctgac ctttggagtt catcaaatgg gatgagccag
     3061 cctggttttg gtggaattct ggggacctcc acttcccaca tgtctcaatc cagtagttat
     3121 ggcaaccttc attcacatga ccgcttgagt tatcctccac actcagtttc accaacagac
     3181 ataaacacga gtcttccacc aatgtccagc tttcatcgcg gcagtaccag cagttcacct
     3241 tacgttgctg cctcacacac tcctcccatc aatggatcag acagcattct aggaaccaga
     3301 gggaatgctg ctggaagctc acagacaggt gatgcacttg gaaaggcttt ggcatctatt
     3361 tattctcctg accataccag cagtagtttt ccgtcaaatc catcaacacc agttggatca
     3421 ccttcacctc tcacaggtac cagtcagtgg ccaagacctg gagggcaagc accttcatcc
     3481 ccaagctatg aaaactcact ccactccctg aaaaatcgag ttgagcagca acttcacgag
     3541 catttgcaag atgcaatgtc cttcttaaag gatgtctgtg agcagtctcg aatggaggat
     3601 cgtttagaca gactggatga tgcaatccat gtgctgcgga accatgctgt gggaccttcc
     3661 accagtttgc ctgctggtca cagtgatata catagtttat tgggaccatc ccataatgca
     3721 ccaattggaa gcctcaattc aaactatgga ggatcaagcc ttgttgcaag cagtcgatca
     3781 gcttcaatgg ttggaactca tcgggaagac tctgtcagtc tcaatggcaa tcattcagtc
     3841 ctgtctagta cagtcactac ttcaagcaca gacctgaacc ataaaacaca agaaaattat
     3901 agaggtggct tgcaaagtca gtctggaact gttgttacaa cagaaatcaa gactgaaaac
     3961 aaagaaaagg atgaaaacct tcatgaacct ccttcatcag atgacatgaa gtcagatgat
     4021 gaatcctccc aaaaagatat caaggtttca tctagaggca gaacaagcag tactaatgaa
     4081 gatgaggatt tgaaccctga acagaagata gaaagggaga aggagaggcg gatggctaac
     4141 aatgccagag aacgcttacg cgtgcgggat attaatgaag cattcaaaga gcttggccga
     4201 atgtgtcagc ttcacttgaa gagtgaaaaa ccccaaacaa aactccttat tcttcatcaa
     4261 gccgtggcag tcatccttag tctagaacag caagtcagag agaggaacct taaccccaaa
     4321 gcagcctgcc ttaagagaag ggaagaagaa aaagtttctg ccgtatcggc agagccgcca
     4381 accacactgc caggaaccca tcctgggctt agtgaaacta ccaaccctat gggtcatatg
     4441 taaggatccg cggccgcaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4501 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4561 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4621 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4681 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4741 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4801 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4861 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4921 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4981 tccaagctgt gaccggcgcc tacgtcgaca tgaaaaagcc tgaactcacc gcgacgtctg
     5041 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     5101 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     5161 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     5221 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     5281 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     5341 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     5401 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     5461 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     5521 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     5581 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     5641 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     5701 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     5761 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     5821 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     5881 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     5941 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     6001 gacgccccag cactcgtccg agggcaaagg aataggtcga caatcaacct ctggattaca
     6061 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     6121 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     6181 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     6241 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     6301 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     6361 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     6421 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     6481 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6541 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     6601 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     6661 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     6721 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6781 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6841 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6901 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     6961 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     7021 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     7081 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     7141 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     7201 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     7261 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     7321 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7381 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     7441 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7501 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7561 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7621 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7681 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7741 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7801 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7861 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7921 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7981 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8041 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8101 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8161 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8221 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8281 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8341 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     8401 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     8461 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8521 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8581 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8641 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8701 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8761 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8821 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8881 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8941 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9001 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9061 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     9121 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9181 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     9241 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     9301 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     9361 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     9421 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     9481 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     9541 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     9601 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     9661 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     9721 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     9781 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     9841 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     9901 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
