LOCUS       V046285                 9921 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046285
VERSION     V046285
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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             1642..5412
                     /label="L1CAM(NM_001278116)"
                     /note="L1CAM(NM_001278116)"
                     /gene="L1CAM"
     polyA_signal    5488..5712
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5758..6186
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6200..6529
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6596..7390
                     /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    7564..7685
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7734..7750)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7758..7774
                     /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(7782..7812)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7827..7848
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8136..8721)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8892..9752)
                     /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(9753..9857)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggtcgtg gcgctgcggt acgtgtggcc tctcctcctc
     1681 tgcagcccct gcctgcttat ccagatcccc gaggaatatg aaggacacca tgtgatggag
     1741 ccacctgtca tcacggaaca gtctccacgg cgcctggttg tcttccccac agatgacatc
     1801 agcctcaagt gtgaggccag tggcaagccc gaagtgcagt tccgctggac gagggatggt
     1861 gtccacttca aacccaagga agagctgggt gtgaccgtgt accagtcgcc ccactctggc
     1921 tccttcacca tcacgggcaa caacagcaac tttgctcaga ggttccaggg catctaccgc
     1981 tgctttgcca gcaataagct gggcaccgcc atgtcccatg agatccggct catggccgag
     2041 ggtgccccca agtggccaaa ggagacagtg aagcccgtgg aggtggagga aggggagtca
     2101 gtggttctgc cttgcaaccc tcccccaagt gcagagcctc tccggatcta ctggatgaac
     2161 agcaagatct tgcacatcaa gcaggacgag cgggtgacga tgggccagaa cggcaacctc
     2221 tactttgcca atgtgctcac ctccgacaac cactcagact acatctgcca cgcccacttc
     2281 ccaggcacca ggaccatcat tcagaaggaa cccattgacc tccgggtcaa ggccaccaac
     2341 agcatgattg acaggaagcc gcgcctgctc ttccccacca actccagcag ccacctggtg
     2401 gccttgcagg ggcagccatt ggtcctggag tgcatcgccg agggctttcc cacgcccacc
     2461 atcaaatggc tgcgccccag tggccccatg ccagccgacc gtgtcaccta ccagaaccac
     2521 aacaagaccc tgcagctgct gaaagtgggc gaggaggatg atggcgagta ccgctgcctg
     2581 gccgagaact cactgggcag tgcccggcat gcgtactatg tcaccgtgga ggctgccccg
     2641 tactggctgc acaagcccca gagccatcta tatgggccag gagagactgc ccgcctggac
     2701 tgccaagtcc agggcaggcc ccaaccagag gtcacctgga gaatcaacgg gatccctgtg
     2761 gaggagctgg ccaaagacca gaagtaccgg attcagcgtg gcgccctgat cctgagcaac
     2821 gtgcagccca gtgacacaat ggtgacccaa tgtgaggccc gcaaccggca cgggctcttg
     2881 ctggccaatg cctacatcta cgttgtccag ctgccagcca agatcctgac tgcggacaat
     2941 cagacgtaca tggctgtcca gggcagcact gcctaccttc tgtgcaaggc cttcggagcg
     3001 cctgtgccca gtgttcagtg gctggacgag gatgggacaa cagtgcttca ggacgaacgc
     3061 ttcttcccct atgccaatgg gaccctgggc attcgagacc tccaggccaa tgacaccgga
     3121 cgctacttct gcctggctgc caatgaccaa aacaatgtta ccatcatggc taacctgaag
     3181 gttaaagatg caactcagat cactcagggg ccccgcagca caatcgagaa gaaaggttcc
     3241 agggtgacct tcacgtgcca ggcctccttt gacccctcct tgcagcccag catcacctgg
     3301 cgtggggacg gtcgagacct ccaggagctt ggggacagtg acaagtactt catagaggat
     3361 gggcgcctgg tcatccacag cctggactac agcgaccagg gcaactacag ctgcgtggcc
     3421 agtaccgaac tggatgtggt ggagagtagg gcacagctct tggtggtggg gagccctggg
     3481 ccggtgccac ggctggtgct gtccgacctg cacctgctga cgcagagcca ggtgcgcgtg
     3541 tcctggagtc ctgcagaaga ccacaatgcc cccattgaga aatatgacat tgaatttgag
     3601 gacaaggaaa tggcgcctga aaaatggtac agtctgggca aggttccagg gaaccagacc
     3661 tctaccaccc tcaagctgtc gccctatgtc cactacacct ttagggttac tgccataaac
     3721 aaatatggcc ccggggagcc cagcccggtc tctgagactg tggtcacacc tgaggcagcc
     3781 ccagagaaga accctgtgga tgtgaagggg gaaggaaatg agaccaccaa tatggtcatc
     3841 acgtggaagc cgctccggtg gatggactgg aacgcccccc aggttcagta ccgcgtgcag
     3901 tggcgccctc aggggacacg agggccctgg caggagcaga ttgtcagcga ccccttcctg
     3961 gtggtgtcca acacgtccac cttcgtgccc tatgagatca aagtccaggc cgtcaacagc
     4021 cagggcaagg gaccagagcc ccaggtcact atcggctact ctggagagga ctacccccag
     4081 gcaatccctg agctggaagg cattgaaatc ctcaactcaa gtgccgtgct ggtcaagtgg
     4141 cggccggtgg acctggccca ggtcaagggc cacctccgcg gatacaatgt gacgtactgg
     4201 agggagggca gtcagaggaa gcacagcaag agacatatcc acaaagacca tgtggtggtg
     4261 cccgccaaca ccaccagtgt catcctcagt ggcttgcggc cctatagctc ctaccacctg
     4321 gaggtgcagg cctttaacgg gcgaggatcg gggcccgcca gcgagttcac cttcagcacc
     4381 ccagagggag tgcctggcca ccccgaggcg ttgcacctgg agtgccagtc gaacaccagc
     4441 ctgctgctgc gctggcagcc cccactcagc cacaacggcg tgctcaccgg ctacgtgctc
     4501 tcctaccacc ccctggatga ggggggcaag gggcaactgt ccttcaacct tcgggacccc
     4561 gaacttcgga cacacaacct gaccgatctc agcccccacc tgcggtaccg cttccagctt
     4621 caggccacca ccaaagaggg ccctggtgaa gccatcgtac gggaaggagg cactatggcc
     4681 ttgtctggga tctcagattt tggcaacatc tcagccacag cgggtgaaaa ctacagtgtc
     4741 gtctcctggg tccccaagga gggccagtgc aacttcaggt tccatatctt gttcaaagcc
     4801 ttgggagaag agaagggtgg ggcttccctt tcgccacagt atgtcagcta caaccagagc
     4861 tcctacacgc agtgggacct gcagcctgac actgactacg agatccactt gtttaaggag
     4921 aggatgttcc ggcaccaaat ggctgtgaag accaatggca caggccgcgt gaggctccct
     4981 cctgctggct tcgccactga gggctggttc atcggctttg tgagtgccat catcctcctg
     5041 ctcctcgtcc tgctcatcct ctgcttcatc aagcgcagca agggcggcaa atactcagtg
     5101 aaggataagg aggacaccca ggtggactct gaggcccgac cgatgaaaga tgagaccttc
     5161 ggcgagtaca ggtccctgga gagtgacaac gaggagaagg cctttggcag cagccagcca
     5221 tcgctcaacg gggacatcaa gcccctgggc agtgacgaca gcctggccga ttatgggggc
     5281 agcgtggatg ttcagttcaa cgaggatggt tcgttcattg gccagtacag tggcaagaag
     5341 gagaaggagg cggcaggggg caatgacagc tcaggggcca cttcccccat caaccctgcc
     5401 gtggccctag aataggatcc tgatatccag cacagtggcg gccgctcgag tctagagggc
     5461 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     5521 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     5581 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     5641 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     5701 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     5761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5941 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     6001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6061 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6181 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     6481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     6541 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     6601 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     6661 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     6721 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     6781 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     6841 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     6901 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     6961 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     7021 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     7081 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     7141 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7201 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7261 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7321 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     7381 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     7441 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7501 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7561 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7621 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7681 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     7741 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7801 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7861 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7921 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7981 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     8041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     8101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8221 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8281 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8341 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8401 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8461 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8521 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     8581 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8641 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     8701 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8761 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8821 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8881 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8941 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     9001 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     9061 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     9121 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     9181 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     9241 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9301 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9361 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9421 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9481 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     9541 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9601 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9661 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9721 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9781 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9841 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     9901 gatctcccga tcccctatgg t
//
