LOCUS       V030493                 9541 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030493
VERSION     V030493
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..3012
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced monomeric blue fluorescent protein
                     (Subach et al., 2011)"
                     /transl_table=1
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
                     /label="mTagBFP2"
     CDS             3043..4194
                     /label="TCF7(NM_003202)"
                     /note="TCF7(NM_003202)"
                     /gene="TCF7"
     CDS             4201..4266
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4310..4521
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4534..4802
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4835..5629
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5636..6224
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6298..6531
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6603..6724
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6743..6878
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6916..6932)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6940..6956
                     /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(6964..6994)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7009..7030
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7318..7906)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8077..8937)
                     /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(8938..9042)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9516..9532
                     /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 catggtgtct aagggcgaag agctgattaa ggagaacatg
     2341 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
     2401 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
     2461 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
     2521 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     2581 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     2641 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
     2701 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc
     2761 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
     2821 ctgatcgcaa acgccaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
     2881 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc
     2941 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
     3001 cacaagctta atggtggagg tggatcagga ggtggaggtt ctatgccgca gctggactcc
     3061 ggcgggggcg gcgcgggcgg cggcgacgac ctcggcgcgc cggacgagct gctggccttc
     3121 caggatgaag gcgaggagca ggacgacaag agccgcgaca gcgccgccgg tcccgagcgc
     3181 gacctggccg agctcaagtc gtcgctcgtg aacgagtccg agggcgcggc cggcggcgca
     3241 gggatcccgg gggtcccggg ggccggcgcc ggggcccgcg gcgaggccga ggctctcggg
     3301 cgggaacacg ctgcgcagag actcttcccg gacaaacttc cagagcccct ggaggacggc
     3361 ctgaaggccc cggagtgcac cagcggcatg tacaaagaga ccgtctactc cgccttcaat
     3421 ctgctcatgc attacccacc cccctcggga gcagggcagc acccccagcc gcagcccccg
     3481 ctgcacaagg ccaatcagcc cccccacggt gtcccccaac tctctctcta cgaacatttc
     3541 aacagcccac atcccacccc tgcacctgcg gacatcagcc agaagcaagt tcacaggcct
     3601 ctgcagaccc ctgacctctc tggcttctac tccctgacct caggcagcat ggggcagctc
     3661 ccccacactg tgagctggtt cacccaccca tccttgatgc taggttctgg tgtacctggt
     3721 cacccagcag ccatccccca cccggccatt gtgcccccct cagggaagca ggagctgcag
     3781 cccttcgacc gcaacctgaa gacacaagca gagtccaagg cagagaagga ggccaagaag
     3841 ccaaccatca agaagcccct caatgccttc atgctgtaca tgaaggagat gagagccaag
     3901 gtcattgcag agtgcacact taaggagagc gctgccatca accagatcct gggccgcagg
     3961 tggcacgcgc tgtcgcgaga agagcaggcc aagtactatg agctggcccg caaggagagg
     4021 cagctgcaca tgcagctata cccaggctgg tcagcgcggg acaactacgg gaagaagaag
     4081 aggcggtcga gggaaaagca ccaagaatcc accacaggag gaaaaagaaa tgcattcggt
     4141 acttacccgg agaaggccgc tgccccagcc ccgttccttc cgatgacagt gctcggatcc
     4201 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4261 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4321 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4381 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4441 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4501 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4561 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4621 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4681 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4741 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4801 tccaagctgt gaccggcgcc tacgctaggc caccatggct gaacaagatg gattgcacgc
     4861 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4921 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4981 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5041 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5101 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5161 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5221 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5281 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5341 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5401 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5461 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5521 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5581 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag tcgacaatca
     5641 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5701 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5761 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5821 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5881 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5941 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6001 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6061 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6121 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6181 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     6241 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6301 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6361 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6421 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6481 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6541 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6601 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6661 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6721 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     6781 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6841 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     6901 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6961 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7021 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7081 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7141 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7201 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7261 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7321 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7381 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7441 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7501 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7561 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7621 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7681 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7741 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7801 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7861 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7921 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7981 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8041 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8101 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8161 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8221 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8281 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8341 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8401 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8461 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8521 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8581 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8641 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8701 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8761 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8821 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8881 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9061 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9121 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9181 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9241 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9301 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9361 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9421 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9481 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     9541 g
//
