LOCUS       V035872                 8500 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035872
VERSION     V035872
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..3354
                     /label="Wnt4(NM_009523)"
                     /note="Wnt4(NM_009523)"
                     /gene="Wnt4"
     CDS             3361..3426
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3470..3681
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3694..3962
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3989..4588
                     /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    4595..5183
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5257..5490
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5562..5683
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5702..5837
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5875..5891)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5899..5915
                     /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(5923..5953)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5968..5989
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6277..6865)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7036..7896)
                     /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(7897..8001)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8475..8491
                     /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 catgagcccc cgttcgtgcc tgcggtccct gcgactcctc
     2341 gtcttcgccg tgttctcggc cgccgcgagc aattggctgt acctggccaa gctgtcatcg
     2401 gtgggcagca tctccgaaga ggagacgtgc gagaaactca aaggcctgat ccagaggcag
     2461 gtgcagatgt gcaaacggaa ccttgaggtg atggactcag tgcgccgtgg tgcccagctg
     2521 gccatcgagg agtgccaata ccagttccgg aaccggcgct ggaactgttc cacactggac
     2581 tccctccctg tctttgggaa ggtggtgaca caagggaccc gggaggcggc ctttgtatac
     2641 gccatctctt cagcaggtgt ggcctttgca gtgacaaggg catgcagcag tggagaactg
     2701 gagaagtgtg gctgtgaccg gacagtgcac ggggtcagcc cacagggctt ccagtggtca
     2761 ggatgctcgg acaacatcgc ctatggcgta gccttctcac agtcctttgt ggacgtccgg
     2821 gagaggagca agggggcctc ctccagccgg gcactcatga atcttcacaa caacgaggct
     2881 ggcaggaagg ccatcttgac acacatgcgg gtggagtgca agtgtcacgg ggtgtcgggc
     2941 tcctgcgagg taaagacgtg ctggcgtgct gtaccgccct tccgccaggt tggccacgcg
     3001 ctaaaggaga agtttgacgg tgccacggag gtggagccac gacgcgtagg ctcctcccgg
     3061 gcgctggtgc ctcggaatgc acagttcaag ccacatacag atgaggacct ggtatacctg
     3121 gagcctagcc cggacttctg tgagcaggac atccgcagtg gcgtgctagg cacgaggggc
     3181 cgcacgtgca acaagacatc taaagccatt gacggctgcg agctactgtg ctgtggccgc
     3241 ggcttccaca cagcgcaagt ggagctggcc gagcgctgtg gctgcaggtt ccactggtgc
     3301 tgcttcgtca agtgccggca gtgccagcgg ctcgtggaga tgcacacgtg ccggggatcc
     3361 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     3421 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     3481 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     3541 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     3601 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     3661 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     3721 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     3781 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     3841 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     3901 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     3961 tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc
     4021 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4081 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4141 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4201 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4261 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4321 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4381 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4441 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4501 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4561 gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg
     4621 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4681 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4741 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4801 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4861 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4921 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4981 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     5041 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5101 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5161 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     5221 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     5281 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     5341 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     5401 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     5461 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     5521 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     5581 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5641 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     5701 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5761 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5821 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     5881 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5941 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6001 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6061 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8041 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8101 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     8161 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     8221 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     8281 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     8341 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8401 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     8461 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
