LOCUS       V033737                 6823 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033737
VERSION     V033737
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..2313
                     /label="Sox3(NM_009237)"
                     /note="Sox3(NM_009237)"
                     /gene="Sox3"
     CDS             2320..2385
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2423..2647
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2693..3121
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3135..3464
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3531..4325
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4499..4620
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4669..4685)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4693..4709
                     /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(4717..4747)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4762..4783
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5071..5656)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5827..6687)
                     /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(6688..6792)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgcgac cagctcgaga gaacgcatca ggtgagagaa gcccgcgagt tcccgccgac
     1021 tttgcgcgga gcccttcggc tagcctgccc ttcccgcctg agctgccggc ccgccggccc
     1081 ctgagcacca ctccgacgga gtcccccggc cttttcacgg tggccgctcc agcccccgga
     1141 gcgccgtctc ctcccgccac gctggcgcac ctccttcccg ccccggcgat gtacagcctg
     1201 ctggagactg aactcaagaa ccccgtgggg ccgcccaccc cagccgcggg caccggcgtc
     1261 cccgcagctc ccggcgctgc aggcaagagt ggcgcgaacc cagccggcgg agcgaacgca
     1321 ggcaacgggg gcagcggggg cgcgaacggc ggcggtggtg gtggtggcgg cgggggcagc
     1381 gaccaggacc gcgtcaagcg acccatgaac gcgttcatgg tgtggtcccg cgggcagcgg
     1441 cgcaagatgg ccctggagaa ccccaagatg cacaactccg agatcagcaa gcgcttgggc
     1501 gccgactgga aactgctgac cgatgcggag aagcggccgt tcatcgacga ggccaagcga
     1561 ctgcgtgcgg tgcacatgaa ggagtacccg gactacaagt accggccccg ccgcaagacc
     1621 aagacgctgc tcaagaagga caagtactcg ctgcccggcg gcctcctgcc cccgggcgcc
     1681 gccgcagccg ccgccgctgc cgccgcagcc gccgccgcca gcagcccggt gggcgtgggc
     1741 cagcgcctgg acacgtacac gcacgtgaac ggctgggcca acggcgccta ctcgctcgtg
     1801 caggagcagc tgggctacgc gcagcccccg agcatgagca gcccgccgcc gccacccgcg
     1861 ctgcctcaga tgcaccgcta cgacatggcc ggcctgcagt acagccccat gatgccaccc
     1921 ggcgcccaga gctacatgaa cgccgccgcc gccgctgccg ccgcctcggg ctacgggggc
     1981 atggcgccct ccgccgcggc cgccgccgcc gccgcctacg ggcagcagcc cgccaccgct
     2041 gccgccgcgg ccgccgccgc cgccgccatg agcctgggcc ccatgggctc cgtggtgaag
     2101 tcggagccca gctctccgcc gcccgccatc gcttcgcact cgcagcgcgc gtgcctcggc
     2161 gacctgcgcg acatgatcag catgtacctg ccacctggcg gggacgcggc cgacgccgct
     2221 tctccgctcc caggcggccg gctgcacggc gtgcaccagc actaccaggg cgccgggact
     2281 gcggtcaatg gaacggtgcc gctgacccac atcctcgagg actacaaaga ccatgacggt
     2341 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgtt
     2401 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2461 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2521 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2581 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2641 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2701 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2761 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2821 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2881 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2941 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3001 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3061 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3121 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3481 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3541 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3601 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3661 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3721 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3781 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3841 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3901 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3961 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4021 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4081 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4141 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4201 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4261 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4321 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4381 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4441 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4501 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4561 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4621 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4681 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4741 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4861 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5581 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     5641 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5701 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5761 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5821 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5881 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5941 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6001 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6061 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6121 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6181 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6241 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6301 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6361 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6421 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6481 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6541 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6601 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6661 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6721 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6781 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
