LOCUS       V035889                 9549 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035889
VERSION     V035889
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..4352
                     /label="CCDC40(NM_017950)"
                     /note="CCDC40(NM_017950)"
                     /gene="CCDC40"
     CDS             4392..5111
                     /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    5149..5373
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5419..5847
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5861..6190
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6257..7051
                     /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    7225..7346
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7395..7411)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7419..7435
                     /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(7443..7473)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7488..7509
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7797..8382)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8553..9413)
                     /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(9414..9518)
                     /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 cggaaccggg cggcgcggcg ggccggtccc
      961 atccggaaga tggatcggct tctgagggag agaaggaagg gaataatgaa agccacatgg
     1021 tgtcaccacc agagaaggat gatggccaga aaggtgaaga agctgtcggt agcacagagc
     1081 atcctgagga agtcacaacc caagcggaag ctgcaattga agagggggag gtggagacag
     1141 aaggggaagc agcagtggaa ggggaagagg aggctgtgtc ctatggagat gctgaaagcg
     1201 aagaggaata ttactataca gaaacttcat ccccggaagg gcaaatcagt gctgcagata
     1261 cgacttaccc gtatttcagt cctcctcagg aactgcctgg agaggaggca tacgatagtg
     1321 ttagcgggga ggctggtctc caaggcttcc agcaagaggc caccggtcca ccagaatcca
     1381 gagaaaggag ggtcacctcc ccagagccat cccacggagt cttaggcccg tcggagcaaa
     1441 tgggccaggt cacctctggg ccagcagtgg gcagattgac aggatccaca gaggagcccc
     1501 aggggcaggt gctcccaatg ggcgtccagc accgcttccg gctgagccac gggagcgaca
     1561 tcgagtcctc agacctggag gagttcgtct cgcaggagcc agtgatcccc ccaggggtgc
     1621 ccgatgccca ccccagggaa ggagacctgc cagtgttcca ggaccagatc cagcagccca
     1681 gcaccgagga gggggccatg gcagagagag tggagtccga ggggagtgac gaggaagcag
     1741 aagacgaagg gtcccagctg gtggttttgg acccagacca ccccctgatg gtaagattcc
     1801 aggctgccct gaagaactac ctgaaccgac agatcgaaaa gttgaagctg gacctccaag
     1861 agctggttgt ggctaccaag cagagccgag cccagcggca ggagctgggg gtgaatctct
     1921 atgaggtgca gcagcacctg gtacacctgc agaagctgct ggagaagagt cacgaccgcc
     1981 acgcaatggc ctcgagcgag cgcaggcaga aggaggagga gctgcaggcc gcccgcgctc
     2041 tctacaccaa gacctgcgca gccgccaacg aggagcgcaa aaagttggcg gctctgcaga
     2101 ctgagatgga gaacttggcc ctgcatctct tctacatgca gaacatcgac caggacatgc
     2161 gtgacgacat ccgcgtgatg acacaagtgg taaagaaggc cgagacggag aggatccggg
     2221 cagaaatcga gaagaaaaag caggacctgt atgtggacca gctcaccact cgagcccagc
     2281 aactggaaga agacattgcc ctgtttgagg ctcagtactt ggcccaagct gaggacaccc
     2341 ggattttaag gaaagcagtg agtgaggcct gcaccgagat cgacgccatc agcgtggaga
     2401 agaggcgcat catgcagcaa tgggccagca gcctggtggg catgaagcac cgcgacgagg
     2461 cgcacagggc ggtgctggag gcgctcagag gatgccagca tcaagccaaa tccaccgacg
     2521 gcgagattga ggcctataag aaatccatca tgaaggagga agaaaagaac gagaagctgg
     2581 cgagcatcct gaaccggaca gagacggaag ccacactgct gcagaagctc accacccagt
     2641 gcctgaccaa gcaggtggcc ctgcagagcc agttcaatac ctacaggctc accctgcagg
     2701 acacagagga tgccctcagc caggaccagc tggaacaaat gatactcacg gaggagttgc
     2761 aggccatccg ccaagccatc cagggcgagc tggagctcag gaggaagacg gatgctgcca
     2821 tccgggagaa gctgcaggag cacatgacct ccaacaagac caccaaatac ttcaaccagc
     2881 tcatcctgag gctgcagaag gagaagacca acatgatgac acatctttcc aaaatcaacg
     2941 gtgacattgc ccagaccacc ctggacatca cacacaccag cagcaggctg gacgcacacc
     3001 agaagaccct ggtggagctg gaccaggacg tgaagaaagt caacgagctc atcaccaaca
     3061 gccagagcga gatctcccgg cgcacgatcc tgatcgagag gaagcaaggg ctcatcaact
     3121 tcctcaacaa gcagctggag cggatggtct ccgagctggg gggggaagaa gtggggcccc
     3181 tggagcttga aatcaaaagg ctgagcaagc tgatcgacga gcacgatggc aaggcggtcc
     3241 aggcccaggt gacctggctg cgcctgcagc aggagatggt caaggtgaca caggagcagg
     3301 aggagcagct ggcctccctg gacgcatcca agaaggagct ccacatcatg gagcagaaga
     3361 aactacgagt agaaagcaag attgagcagg agaagaagga gcagaaggag atcgagcacc
     3421 acatgaagga cctggacaac gacctgaaga agctcaacat gttgatgaat aaaaaccggt
     3481 gcagctcgga ggagctggag cagaacaacc gggtgacaga gaatgagttc gtgcgctcgc
     3541 tgaaggcctc tgagagggag accatcaaga tgcaggacaa gctgaaccag ctcagcgagg
     3601 agaaggcgac cctcctgaat caactggtgg aagcagaaca ccagattatg ctttgggaga
     3661 aaaaaatcca actggcaaaa gagatgcgtt cctcagtgga ttccgagatc ggccagacgg
     3721 agatccgggc catgaagggc gagatccaca ggatgaaggt caggctcggg cagctgctga
     3781 agcagcagga gaagatgatc cgtgccatgg agttggcggt tgcccgcaga gagaccgtca
     3841 ccacccaggc cgaggggcag cgcaagatgg acaggaaggc gctcacccgc accgacttcc
     3901 accacaagca gcttgagctg cgccggaaaa tcagggacgt tcgcaaggcc accgatgagt
     3961 gcaccaaaac cgtcctggaa ctggaagaaa cacaaagaaa tgtgagcagc tccctcctag
     4021 agaagcagga aaagctgtcg gtgattcagg cagacttcga cacactcgag gccgacctca
     4081 cccggcttgg ggccctcaaa cgacagaacc tttcagagat cgtggccctg cagacacgcc
     4141 ttaagcacct gcaggctgtg aaggaggggc gctacgtgtt cctgttccgc tccaagcagt
     4201 ccctagtgct ggagcgccag cgcctggaca agcgactggc tctcatcgcc accatcctgg
     4261 accgcgtgcg ggacgagtac ccccagttcc aggaggccct gcacaaggtc agccagatga
     4321 tcgccaacaa gctcgagtca ccagggccct cctcctcgag tggcggtgga ggaagcggcg
     4381 gaggtggatc tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     4441 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     4501 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     4561 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     4621 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     4681 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     4741 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     4801 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     4861 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     4921 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     4981 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     5041 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     5101 tgtacaagta gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     5161 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5221 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5281 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     5341 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     5401 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5461 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5521 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5581 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5641 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5701 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5761 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5821 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     5881 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5941 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6001 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6061 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6121 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6181 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     6241 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     6301 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     6361 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     6421 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     6481 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     6541 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     6601 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     6661 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     6721 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6781 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6841 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6901 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6961 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     7021 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     7081 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     7141 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     7201 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     7261 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     7321 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     7381 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7441 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7501 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7561 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7621 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7681 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7741 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7801 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7861 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7921 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7981 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8041 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8101 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8161 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8221 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8281 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt
     8341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8521 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8581 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8641 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8701 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8761 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8821 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8881 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8941 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     9001 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     9061 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     9121 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     9181 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     9241 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     9301 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     9361 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     9421 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9481 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9541 cctgacgtc
//
