LOCUS       V029369                 5989 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029369
VERSION     V029369
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..1479
                     /label="CRIPTO(NM_001174136)"
                     /note="CRIPTO(NM_001174136)"
                     /gene="CRIPTO"
     CDS             1486..1551
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1589..1813
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      1859..2287
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2301..2630
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2697..3491
                     /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    3665..3786
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3835..3851)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3859..3875
                     /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(3883..3913)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3928..3949
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4237..4822)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4993..5853)
                     /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(5854..5958)
                     /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 accatggcca tttctaaagt ctttgaactg ggattagttg ccgggctggg ccatcaggaa
     1021 tttgctcgtc catctcgggg atacctggcc ttcagagatg acagcatttg gccccaggag
     1081 gagcctgcaa ttcggcctcg gtcttcccag cgtgtgccgc ccatggggat acagcacagt
     1141 aaggagctaa acagaacctg ctgcctgaat gggggaacct gcatgctggg gtccttttgt
     1201 gcctgccctc cctccttcta cggacggaac tgtgagcacg atgtgcgcaa agagaactgt
     1261 gggtctgtgc cccatgacac ctggctgccc aagaagtgtt ccctgtgtaa atgctggcac
     1321 ggtcagctcc gctgctttcc tcaggcattt ctacccggct gtgatggcct tgtgatggat
     1381 gagcacctcg tggcttccag gactccagaa ctaccaccgt ctgcacgtac taccactttt
     1441 atgctagttg gcatctgcct ttctatacaa agctactatc tcgaggacta caaagaccat
     1501 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg
     1561 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1621 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1681 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1741 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1801 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1861 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1921 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1981 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2041 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2101 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2161 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2221 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2281 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2341 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2401 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2461 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2521 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2581 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2641 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2701 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2761 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2821 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     2881 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2941 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3001 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3061 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3121 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3181 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3241 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3301 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3361 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3421 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3481 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3541 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3601 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3661 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3781 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3841 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3901 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4741 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     4801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4861 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4921 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4981 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5041 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5101 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5161 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5221 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5281 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5341 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5401 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5461 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5521 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5581 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5641 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5701 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5761 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5821 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5881 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5941 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
