LOCUS       V032243                 7629 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032243
VERSION     V032243
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             919..2442
                     /label="CREB5(NM_182898)"
                     /note="CREB5(NM_182898)"
                     /gene="CREB5"
     CDS             2479..3198
                     /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    3229..3453
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3499..3927
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3941..4270
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4337..5131
                     /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    5305..5426
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5475..5491)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5499..5515
                     /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(5523..5553)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5568..5589
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5877..6462)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6633..7493)
                     /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(7494..7598)
                     /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 gatttatgag gaatccaaga tgaatttgga gcaggagagg
      961 ccgtttgtct gcagtgcccc aggctgctcc cagcgcttcc caacagagga ccatctgatg
     1021 attcataggc acaaacatga aatgactttg aagtttcctt caataaaaac agacaatatg
     1081 ttatcagatc aaactccgac cccaacgaga ttcctgaaga actgcgagga ggtgggcctc
     1141 ttcagcgagc tggactgctc cctggagcac gagttcagga aggctcagga agaggagagc
     1201 agcaagcgga atatctcgat gcataatgca gttggtgggg ccatgacggg gcccggaact
     1261 caccagctta gcagcgctcg gctgcccaac catgacacca acgttgtgat tcagcaagcc
     1321 atgccgtcgc ctcagtccag ctctgtcatc actcaggcac cttccaccaa ccgccagatc
     1381 gggcctgtcc caggctctct atcttctctg ctacatctcc acaacagaca gagacagccc
     1441 atgccagcct ccatgcctgg gaccctgccc aaccctacaa tgccaggatc ttccgccgtc
     1501 ttgatgccaa tggagcgaca aatgtcagtg aactccagca tcatggggat gcaaggtcca
     1561 aatctcagca acccctgtgc ttctccccag gtccagccaa tgcattcaga agccaaaatg
     1621 aggttgaagg ctgcattgac tcaccaccct gctgccatgt caaatgggaa catgaacacc
     1681 atgggacaca tgatggagat gatgggctcc cggcaggacc agacgccaca ccatcacatg
     1741 cactcgcacc cgcatcagca ccagacactg ccaccccatc acccttaccc acaccagcac
     1801 cagcacccag cacaccatcc tcaccctcaa ccccatcacc agcagaacca tccacatcac
     1861 cactcccatt cccaccttca tgcacaccca gcacatcacc agacctcgcc acatccgccc
     1921 ctgcacaccg gcaaccaagc acaggtttca ccagcaacac aacagatgca gccaacccag
     1981 acaatacagc caccccagcc cacagggggg cgccggcgaa gggtggtaga cgaggatccg
     2041 gacgagaggc ggcggaaatt tctggaacgg aaccgggcag ctgccacccg ctgcagacag
     2101 aagaggaagg tctgggtgat gtcattggaa aagaaagcag aagaactcac ccagacaaac
     2161 atgcagcttc agaatgaagt gtctatgttg aaaaatgagg tggcccagct gaaacagttg
     2221 ttgttaacac ataaagactg cccaataaca gccatgcaga aagaatcaca aggatatcta
     2281 agtccagaga gtagccctcc tgctagtcct gtcccagctt gctcccagca acaagtcatc
     2341 cagcataata ccatcactac ttcctcatcg gtcagcgagg tggtaggaag ctccaccctc
     2401 agccagctca ccactcacag aacagacctg aatccgattc tttcgagtgg cggtggagga
     2461 agcggcggag gtggatctat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2521 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2581 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2641 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2701 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2761 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2821 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2881 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2941 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3001 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3061 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3121 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3181 gacgagctgt acaagtaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     3241 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3301 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3361 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3421 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3481 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3541 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3601 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3661 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3721 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3781 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3841 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3901 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3961 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4021 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4081 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4141 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4201 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4261 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4321 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4381 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4441 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4501 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4561 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4621 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4681 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4741 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4801 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4861 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4921 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4981 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5041 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5101 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5161 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5221 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5281 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5341 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5401 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5461 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5521 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5581 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6301 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt
     6421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6481 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6541 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6601 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6661 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6721 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6781 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6841 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6901 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6961 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7021 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7081 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7141 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7201 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7261 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7321 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7381 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7441 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7501 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7561 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7621 cctgacgtc
//
