LOCUS       V023211                 7181 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023211
VERSION     V023211
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             941..2671
                     /label="IGF2BP1(NM_006546)"
                     /note="IGF2BP1(NM_006546)"
                     /gene="IGF2BP1"
     CDS             2678..2743
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2781..3005
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3051..3479
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3493..3822
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3889..4683
                     /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    4857..4978
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5027..5043)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5051..5067
                     /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(5075..5105)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5120..5141
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5429..6014)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6185..7045)
                     /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(7046..7150)
                     /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 atccgccacc atgaacaagc tttacatcgg
      961 caacctcaac gagagcgtga cccccgcgga cttggagaaa gtgtttgcgg agcacaagat
     1021 ctcctacagc ggccagttct tggtcaaatc cggctacgcc ttcgtggact gcccggacga
     1081 gcactgggcg atgaaggcca tcgaaacttt ctccgggaaa gtagaattac aaggaaaacg
     1141 cttagagatt gaacattcgg tgcccaaaaa acaaaggagc cggaaaattc aaatccgaaa
     1201 tattccaccc cagctccgat gggaagtact ggacagcctg ctggctcagt atggtacagt
     1261 agagaactgt gagcaagtga acaccgagag tgagacggca gtggtgaatg tcacctattc
     1321 caaccgggag cagaccaggc aagccatcat gaagctgaat ggccaccagt tggagaacca
     1381 tgccctgaag gtctcctaca tccccgatga gcagatagca cagggacctg agaatgggcg
     1441 ccgagggggc tttggctctc ggggtcagcc ccgccagggc tcacctgtgg cagcgggggc
     1501 cccagccaag cagcagcaag tggacatccc ccttcggctc ctggtgccca cccagtatgt
     1561 gggtgccatt attggcaagg agggggccac catccgcaac atcacaaaac agacccagtc
     1621 caagatagac gtgcatagga aggagaacgc aggtgcagct gaaaaagcca tcagtgtgca
     1681 ctccacccct gagggctgct cctccgcttg taagatgatc ttggagatta tgcataaaga
     1741 ggctaaggac accaaaacgg ctgacgaggt tcccctgaag atcctggccc ataataactt
     1801 tgtagggcgt ctcattggca aggaaggacg gaacctgaag aaggtagagc aagataccga
     1861 gacaaaaatc accatctcct cgttgcaaga ccttaccctt tacaaccctg agaggaccat
     1921 cactgtgaag ggggccatcg agaattgttg cagggccgag caggaaataa tgaagaaagt
     1981 tcgggaggcc tatgagaatg atgtggctgc catgagcctg cagtctcacc tgatccctgg
     2041 cctgaacctg gctgctgtag gtcttttccc agcttcatcc agcgcagtcc cgccgcctcc
     2101 cagcagcgtt actggggctg ctccctatag ctcctttatg caggctcccg agcaggagat
     2161 ggtgcaggtg tttatccccg cccaggcagt gggcgccatc atcggcaaga aggggcagca
     2221 catcaaacag ctctcccggt ttgccagcgc ctccatcaag attgcaccac ccgaaacacc
     2281 tgactccaaa gttcgtatgg ttatcatcac tggaccgcca gaggcccaat tcaaggctca
     2341 gggaagaatc tatggcaaac tcaaggagga gaacttcttt ggtcccaagg aggaagtgaa
     2401 gctggagacc cacatacgtg tgccagcatc agcagctggc cgggtcattg gcaaaggtgg
     2461 aaaaacggtg aacgagttgc agaatttgac ggcagctgag gtggtagtac caagagacca
     2521 gacccctgat gagaacgacc aggtcatcgt gaaaatcatc ggacatttct atgccagtca
     2581 gatggctcaa cggaagatcc gagacatcct ggcccaggtt aagcagcagc atcagaaggg
     2641 acagagtaac caggcccagg cacggaggaa gctcgaggac tacaaagacc atgacggtga
     2701 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     2761 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2821 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2881 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2941 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3001 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3061 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3121 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3181 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3241 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3301 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3361 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3421 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3481 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3721 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3841 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3901 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3961 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4021 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4081 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4141 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4201 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4261 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4321 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4381 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4441 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4501 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4561 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4621 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4681 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4741 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4801 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4861 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4921 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4981 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5041 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5101 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5161 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5221 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5281 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5341 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5401 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5461 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5521 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5581 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5641 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5701 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5761 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5821 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5881 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5941 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7141 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
