LOCUS       V022114                 8410 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022114
VERSION     V022114
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             649..1365
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1399..3135
                     /label="IGF2BP3(NM_006547)"
                     /note="IGF2BP3(NM_006547)"
                     /gene="IGF2BP3"
     CDS             3136..3153
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     misc_feature    3186..3774
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3840..3888
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4090..4518
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4532..4861
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4928..5722
                     /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    5898..6019
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6068..6084)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6092..6108
                     /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(6116..6146)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6161..6182
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6470..7058)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7229..8089)
                     /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(8090..8194)
                     /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 gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccgagc tcggatccat gaacaaactg tatatcggaa acctcagcga gaacgccgcc
     1441 ccctcggacc tagaaagtat cttcaaggac gccaagatcc cggtgtcggg acccttcctg
     1501 gtgaagactg gctacgcgtt cgtggactgc ccggacgaga gctgggccct caaggccatc
     1561 gaggcgcttt caggtaaaat agaactgcac gggaaaccca tagaagttga gcactcggtc
     1621 ccaaaaaggc aaaggattcg gaaacttcag atacgaaata tcccgcctca tttacagtgg
     1681 gaggtgctgg atagtttact agtccagtat ggagtggtgg agagctgtga gcaagtgaac
     1741 actgactcgg aaactgcagt tgtaaatgta acctattcca gtaaggacca agctagacaa
     1801 gcactagaca aactgaatgg atttcagtta gagaatttca ccttgaaagt agcctatatc
     1861 cctgatgaaa tggccgccca gcaaaacccc ttgcagcagc cccgaggtcg ccgggggctt
     1921 gggcagaggg gctcctcaag gcaggggtct ccaggatccg tatccaagca gaaaccatgt
     1981 gatttgcctc tgcgcctgct ggttcccacc caatttgttg gagccatcat aggaaaagaa
     2041 ggtgccacca ttcggaacat caccaaacag acccagtcta aaatcgatgt ccaccgtaaa
     2101 gaaaatgcgg gggctgctga gaagtcgatt actatcctct ctactcctga aggcacctct
     2161 gcggcttgta agtctattct ggagattatg cataaggaag ctcaagatat aaaattcaca
     2221 gaagagatcc ccttgaagat tttagctcat aataactttg ttggacgtct tattggtaaa
     2281 gaaggaagaa atcttaaaaa aattgagcaa gacacagaca ctaaaatcac gatatctcca
     2341 ttgcaggaat tgacgctgta taatccagaa cgcactatta cagttaaagg caatgttgag
     2401 acatgtgcca aagctgagga ggagatcatg aagaaaatca gggagtctta tgaaaatgat
     2461 attgcttcta tgaatcttca agcacattta attcctggat taaatctgaa cgccttgggt
     2521 ctgttcccac ccacttcagg gatgccacct cccacctcag ggcccccttc agccatgact
     2581 cctccctacc cgcagtttga gcaatcagaa acggagactg ttcatctgtt tatcccagct
     2641 ctatcagtcg gtgccatcat cggcaagcag ggccagcaca tcaagcagct ttctcgcttt
     2701 gctggagctt caattaagat tgctccagcg gaagcaccag atgctaaagt gaggatggtg
     2761 attatcactg gaccaccaga ggctcagttc aaggctcagg gaagaattta tggaaaaatt
     2821 aaagaagaaa actttgttag tcctaaagaa gaggtgaaac ttgaagctca tatcagagtg
     2881 ccatcctttg ctgctggcag agttattgga aaaggaggca aaacggtgaa tgaacttcag
     2941 aatttgtcaa gtgcagaagt tgttgtccct cgtgaccaga cacctgatga gaatgaccaa
     3001 gtggttgtca aaataactgg tcacttctat gcttgccagg ttgcccagag aaaaattcag
     3061 gaaattctga ctcaggtaaa gcagcaccaa caacagaagg ctctgcaaag tggaccacct
     3121 cagtcaagac ggaagcacca tcaccatcac cattagctcg agtctagagg gcccgatatc
     3181 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3241 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3301 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3361 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3421 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3481 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3541 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3601 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3661 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3721 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     3781 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     3841 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     3901 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3961 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4021 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     4081 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4141 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4201 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     4261 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4321 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4381 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4441 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     4501 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4561 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4621 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4681 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4741 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4801 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4861 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4921 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4981 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5041 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5101 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5161 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5221 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5281 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5341 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5401 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5461 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5521 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5581 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5641 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5701 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     5761 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5821 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5881 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5941 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6001 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     6061 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6121 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6181 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6301 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6361 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6661 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6901 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8221 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     8281 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     8341 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     8401 tctgcttagg
//
