LOCUS       V039418                 8139 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039418
VERSION     V039418
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             927..2942
                     /label="SHCBP1(NM_024745)"
                     /note="SHCBP1(NM_024745)"
                     /gene="SHCBP1"
     CDS             2982..3701
                     /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"
     polyA_signal    3739..3963
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4009..4437
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4451..4780
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4847..5641
                     /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    5815..5936
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5985..6001)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6009..6025
                     /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(6033..6063)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6078..6099
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6387..6972)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7143..8003)
                     /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(8004..8108)
                     /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 gtttaaagcc actgggatcc gccaccatgg ctgacgggtc gctgacgggc ggcggtctgg
      961 aggcagcggc catggcgccg gagcgcatgg gctgggcggt ggagcaggag ctggcgtctc
     1021 tggagaaagg tttgttccaa gatgaagatt catgcagtga ttgtagctac cgtgataaac
     1081 caggttctag tttacaaagt tttatgccag aaggaaaaac ctttttccca gaaattttcc
     1141 aaacaaatca acttttgttc tatgagcgat tcagagccta tcaagattac attttagctg
     1201 actgcaaggc ctctgaggta caggaattca cagctgagtt cttggagaag gtccttgagc
     1261 catctggatg gcgggcagtc tggcacacta atgtgttcaa ggtgctggtt gagatcacag
     1321 atgtggactt tgcagccttg aaggcagtgg tgaggcttgc tgaaccatac ctctgtgact
     1381 ctcaagtgag cacttttacc atggagtgca tgaaggagct ccttgatctg aaggagcatc
     1441 ggttgcccct gcaggagctg tgggtggtgt ttgatgattc aggagtgttt gaccagacag
     1501 cccttgcaat tgagcatgtc agatttttct accaaaacat ttggaggagt tgggatgaag
     1561 aagaggagga tgaatacgat tattttgtca gatgtgttga acctcgatta agattgcatt
     1621 atgacattct tgaagaccga gttccatcag gacttattgt tgactaccac aatctgttgt
     1681 ctcaatgtga ggagagttac aggaaatttt taaatctgag aagcagtttg tcaaattgta
     1741 actctgattc cgagcaggaa aatatctcca tggtggaagg gttaaaattg tattcggaga
     1801 tggaacagtt gaaacaaaag ctgaaactca ttgagaatcc tttgttgagg tatgtgtttg
     1861 gttatcagaa gaattctaac atccaagcaa agggtgtccg ttccagcggt cagaagatca
     1921 ctcatgtggt ctcctccacc atgatggctg gtctcctgcg gtccctgctt acggacaggc
     1981 tttgccagga gcctggtgag gaagaaagag aaattcagtt ccatagtgat ccattgtctg
     2041 ctataaatgc ctgcttcgaa ggtgacactg ttattgtttg tcctggccat tatgtggtac
     2101 atggcacttt ctccattgct gactccattg agttggaagg atatggccta ccagatgaca
     2161 ttgtgataga aaagaggggc aaaggcgaca cttttgtgga ctgcactggt gctgatatta
     2221 aaatctcagg cataaaattt gttcagcatg atgctgtaga gggaatctta attgttcacc
     2281 gtggtaagac tacgctggaa aactgtgtgc tgcagtgtga gacgaccgga gtcacagtgc
     2341 ggacatcagc agagtttcta atgaagaact cggatttata tggcgccaag ggtgctggta
     2401 tagaaatcta ccctgggagt cagtgcaccc tgagtgacaa tgggatccat cactgcaagg
     2461 aagggatcct cattaaggac ttcttagatg aacattatga cattcccaag atatccatgg
     2521 tgaataatat aatacataat aatgaaggtt atggtgttgt cttggtgaaa cctacaatct
     2581 tctctgacct gcaagaaaat gctgaagatg gaactgaaga aaataaagcg cttaaaattc
     2641 agacaagtgg agagccagat gtggctgaaa gagtggatct agaagagctg attgagtgtg
     2701 caactggtaa aatggagctt tgtgcaagaa ctgacccttc tgagcaagtc gagggaaatt
     2761 gtgaaattgt aaatgaacta attgctgcct ccacacagaa aggccagata aagaagaaaa
     2821 ggttgagtga actggggatc acgcaagctg atgacaactt aatgtcacag gagatgtttg
     2881 ttgggattgt ggggaaccag ttcaagtgga atgggaaagg tagttttggc acatttcttt
     2941 tctcctcgag tggcggtgga ggaagcggcg gaggtggatc tatggtgagc aagggcgagg
     3001 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3061 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3121 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3181 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3241 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3301 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3361 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3421 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3481 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3541 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3601 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3661 ccgccgggat cactctcggc atggacgagc tgtacaagta gtctagaggg cccgtttaaa
     3721 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3781 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3841 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3901 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3961 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     4021 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4081 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4141 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4201 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4261 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4321 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4381 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4441 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4501 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4561 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4621 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4681 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4741 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4801 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4861 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4921 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4981 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5041 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5101 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5161 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5221 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5281 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5341 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5401 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5461 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5521 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5581 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5641 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5701 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5761 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5821 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5881 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5941 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6001 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6061 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6121 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6181 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6901 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6961 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7021 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7081 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7141 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7201 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7261 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7321 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7381 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7441 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7501 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7561 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7621 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7681 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7741 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7801 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7861 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7921 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7981 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8101 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
