LOCUS       V037046                 6779 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037046
VERSION     V037046
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..1459
                     /label="IGFBP1(NM_000596)"
                     /note="IGFBP1(NM_000596)"
                     /gene="IGFBP1"
     CDS             1466..1531
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1555..2143
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2209..2257
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2459..2887
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2901..3230
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3297..4091
                     /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    4267..4388
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4437..4453)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4461..4477
                     /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(4485..4515)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4530..4551
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4839..5427)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5598..6458)
                     /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(6459..6563)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgtcaga ggtccccgtt gctcgcgtct ggctggtact
      721 gctcctgctg actgtccagg tcggcgtgac agccggcgct ccgtggcagt gcgcgccctg
      781 ctccgccgag aagctcgcgc tctgcccgcc ggtgtccgcc tcgtgctcgg aggtcacccg
      841 gtccgccggc tgcggctgtt gcccgatgtg cgccctgcct ctgggcgccg cgtgcggcgt
      901 ggcgactgca cgctgcgccc ggggactcag ttgccgcgcg ctgccggggg agcagcaacc
      961 tctgcacgcc ctcacccgcg gccaaggcgc ctgcgtgcag gagtctgacg cctccgctcc
     1021 ccatgctgca gaggcaggga gccctgaaag cccagagagc acggagataa ctgaggagga
     1081 gctcctggat aatttccatc tgatggcccc ttctgaagag gatcattcca tcctttggga
     1141 cgccatcagt acctatgatg gctcgaaggc tctccatgtc accaacatca aaaaatggaa
     1201 ggagccctgc cgaatagaac tctacagagt cgtagagagt ttagccaagg cacaggagac
     1261 atcaggagaa gaaatttcca aattttacct gccaaactgc aacaagaatg gattttatca
     1321 cagcagacag tgtgagacat ccatggatgg agaggcggga ctctgctggt gcgtctaccc
     1381 ttggaatggg aagaggatcc ctgggtctcc agagatcagg ggagacccca actgccagat
     1441 atattttaat gtacaaaacc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1501 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     1561 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1621 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1681 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     1741 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     1801 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     1861 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1921 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     1981 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2041 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2101 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2161 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2221 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2281 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2341 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2401 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2581 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2821 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2881 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3241 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3301 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3361 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3421 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3481 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3541 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3601 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3661 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3721 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3781 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3841 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3901 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3961 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4021 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4081 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4141 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4201 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4261 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4321 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4381 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4441 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4501 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4561 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6481 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6541 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6601 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     6661 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     6721 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
