LOCUS       V044235                 7203 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044235
VERSION     V044235
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2721
                     /label="Igf2bp1(NM_009951)"
                     /note="Igf2bp1(NM_009951)"
                     /gene="Igf2bp1"
     polyA_signal    2800..3024
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3070..3498
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3512..3841
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3908..4702
                     /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    4876..4997
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5046..5062)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5070..5086
                     /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(5094..5124)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5139..5160
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5448..6036)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6207..7067)
                     /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(7068..7172)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgaacaagc tttacatcgg caacctcaac
     1021 gagagtgtga cccccgcaga cttggagaaa gtattcgcgg agcacaagat ctcctacagc
     1081 ggccagttct tggtcaaatc cggctacgcc ttcgtggatt gccccgacga gcactgggcg
     1141 atgaaggcca tcgaaacttt ctcggggaaa gtagaactgc aaggaaaacg tctagagatt
     1201 gaacactcag tccccaaaaa acaaaggagt cggaaaatac agatccgcaa tattccacct
     1261 cagctccgat gggaagtgct agatagcctg ctggctcagt acggtacagt ggagaactgt
     1321 gagcaagtga acactgaaag tgagacagct gtggtcaacg tcacctactc taaccgggag
     1381 cagaccaggc aagctatcat gaagctaaat ggccatcaac tggagaacca tgccctgaag
     1441 gtctcctaca tacctgatga gcagataacg caaggtcctg agaatgggcg tcgtggaggc
     1501 tttgggtctc ggggccagcc ccggcaaggg tcgcccgtgg cagcaggggc tccagccaag
     1561 cagcagccag tggacatccc tctccggctc ctggtgccta cgcagtatgt aggcgctatc
     1621 attggcaagg agggtgccac catccgaaac atcacaaaac agacgcagtc caaaatagac
     1681 gtgcatagga aggagaatgc gggcgctgcg gagaaggcca tcagcgtgca ttcaacccct
     1741 gaaggctgct cctccgcgtg caagatgatc ttggagatta tgcacaagga ggcaaaggac
     1801 accaaaacgg cagatgaagt tcccctgaag atcctggctc ataacaactt cgtcgggcga
     1861 ctcattggca aggaagggcg gaacctgaag aaggtggagc aggacacaga gacgaagatc
     1921 accatctcat cgctccagga cctcacgctc tataaccctg agaggaccat cactgtgaag
     1981 ggcgccattg agaactgttg cagggccgag caggagatca tgaagaaagt tcgagaggct
     2041 tacgagaacg acgtggccgc catgagcttg cagtcccacc tcatccctgg gcttaacctg
     2101 gctgctgtag gtctcttccc agcttcatcc agcgctgtcc ctcctcctcc cagcagtgtc
     2161 accggggctg ctccctatag ctccttcatg caggctccgg agcaggagat ggtacaagtg
     2221 ttcatccccg cccaggctgt gggcgccatc attggcaaga agggccagca catcaaacaa
     2281 ctctcccgct tcgccagcgc ctccatcaag attgcaccac cagaaacacc tgactccaaa
     2341 gttcgaatgg tcgtcatcac tggaccccca gaggctcagt tcaaggccca gggaagaatc
     2401 tatggcaaac taaaagaaga gaatttcttt ggtcccaagg aggaagtaaa gctagagacc
     2461 cacatacggg ttccggcttc agcagccggc cgtgtcatcg gcaaaggcgg caaaacggtg
     2521 aatgagctgc agaacttgac cgcagctgag gtggtagtgc caagagacca gaccccggat
     2581 gagaacgacc aagtcattgt taagatcatc ggacatttct atgccagcca gatggctcag
     2641 cggaagatcc gagacatcct ggctcaagtt aagcaacagc accagaaggg acagagcaac
     2701 ctggcccagg ctcggaggaa gtagaattct gcagatatcc agcacagtgg cggccgctcg
     2761 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     2821 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2881 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2941 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3001 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3061 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3121 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3181 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3241 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3301 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3361 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3421 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3481 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3541 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3661 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3721 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3781 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3841 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3901 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3961 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4021 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4081 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4141 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4201 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4261 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4321 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4381 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4441 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4501 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4561 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4621 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4681 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4741 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4801 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4861 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4921 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4981 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5041 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5101 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5881 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7201 gtc
//
