LOCUS       V038577                 9149 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038577
VERSION     V038577
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             923..3958
                     /label="KL(NM_004795)"
                     /note="KL(NM_004795)"
                     /gene="KL"
     CDS             4001..4714
                     /codon_start=1
                     /note="excitable with blue light"
                     /product="S65T variant of Aequorea victoria green
                     fluorescent protein (Heim et al., 1995)"
                     /transl_table=1
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
                     /label="GFP (S65T)"
     polyA_signal    4749..4973
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5019..5447
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5461..5790
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5857..6651
                     /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    6825..6946
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6995..7011)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7019..7035
                     /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(7043..7073)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7088..7109
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7397..7982)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8153..9013)
                     /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(9014..9118)
                     /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 aagcttgcca ccatgcccgc cagcgccccg ccgcgccgcc cgcggccgcc
      961 gccgccgtcg ctgtcgctgc tgctggtgct gctgggcctg ggcggccgcc gcctgcgtgc
     1021 ggagccgggc gacggcgcgc agacctgggc ccgtttctcg cggcctcctg cccccgaggc
     1081 cgcgggcctc ttccagggca ccttccccga cggcttcctc tgggccgtgg gcagcgccgc
     1141 ctaccagacc gagggcggct ggcagcagca cggcaagggt gcgtccatct gggatacgtt
     1201 cacccaccac cccctggcac ccccgggaga ctcccggaac gccagtctgc cgttgggcgc
     1261 cccgtcgccg ctgcagcccg ccaccgggga cgtagccagc gacagctaca acaacgtctt
     1321 ccgcgacacg gaggcgctgc gcgagctcgg ggtcactcac taccgcttct ccatctcgtg
     1381 ggcgcgagtg ctccccaatg gcagcgcggg cgtccccaac cgcgaggggc tgcgctacta
     1441 ccggcgcctg ctggagcggc tgcgggagct gggcgtgcag cccgtggtca ccctgtacca
     1501 ctgggacctg ccccagcgcc tgcaggacgc ctacggcggc tgggccaacc gcgccctggc
     1561 cgaccacttc agggattacg cggagctctg cttccgccac ttcggcggtc aggtcaagta
     1621 ctggatcacc atcgacaacc cctacgtggt ggcctggcac ggctacgcca ccgggcgcct
     1681 ggcccccggc atccggggca gcccgcggct cgggtacctg gtggcgcaca acctcctcct
     1741 ggctcatgcc aaagtctggc atctctacaa tacttctttc cgtcccactc agggaggtca
     1801 ggtgtccatt gccctaagct ctcactggat caatcctcga agaatgaccg accacagcat
     1861 caaagaatgt caaaaatctc tggactttgt actaggttgg tttgccaaac ccgtatttat
     1921 tgatggtgac tatcccgaga gcatgaagaa taacctttca tctattctgc ctgattttac
     1981 tgaatctgag aaaaagttca tcaaaggaac tgctgacttt tttgctcttt gctttggacc
     2041 caccttgagt tttcaacttt tggaccctca catgaagttc cgccaattgg aatctcccaa
     2101 cctgaggcaa ctgctttcct ggattgacct tgaatttaac catcctcaaa tatttattgt
     2161 ggaaaatggc tggtttgtct cagggaccac caagagagat gatgccaaat atatgtatta
     2221 cctcaaaaag ttcatcatgg aaaccttaaa agccatcaag ctggatgggg tggatgtcat
     2281 cgggtatacc gcatggtccc tcatggatgg tttcgagtgg cacagaggtt acagcatcag
     2341 gcgtggactc ttctatgttg actttctaag ccaggacaag atgttgttgc caaagtcttc
     2401 agccttgttc taccaaaagc tgatagagaa aaatggcttc cctcctttac ctgaaaatca
     2461 gcccctagaa gggacatttc cctgtgactt tgcttgggga gttgttgaca actacattca
     2521 agtagatacc actctgtctc agtttaccga cctgaatgtt tacctgtggg atgtccacca
     2581 cagtaaaagg cttattaaag tggatggggt tgtgaccaag aagaggaaat cctactgtgt
     2641 tgactttgct gccatccagc cccagatcgc tttactccag gaaatgcacg ttacacattt
     2701 tcgcttctcc ctggactggg ccctgattct ccctctgggt aaccagtccc aggtgaacca
     2761 caccatcctg cagtactatc gctgcatggc cagcgagctt gtccgtgtca acatcacccc
     2821 agtggtggcc ctgtggcagc ctatggcccc gaaccaagga ctgccgcgcc tcctggccag
     2881 gcagggcgcc tgggagaacc cctacactgc cctggccttt gcagagtatg cccgactgtg
     2941 ctttcaagag ctcggccatc acgtcaagct ttggataacg atgaatgagc cgtatacaag
     3001 gaatatgaca tacagtgctg gccacaacct tctgaaggcc catgccctgg cttggcatgt
     3061 gtacaatgaa aagtttaggc atgctcagaa tgggaaaata tccatagcct tgcaggctga
     3121 ttggatagaa cctgcctgcc ctttctccca aaaggacaaa gaggtggctg agagagtttt
     3181 ggaatttgac attggctggc tggctgagcc cattttcggc tctggagatt atccatgggt
     3241 gatgagggac tggctgaacc aaagaaacaa ttttcttctt ccttatttca ctgaagatga
     3301 aaaaaagcta atccagggta cctttgactt tttggcttta agccattata ccaccatcct
     3361 tgtagactca gaaaaagaag atccaataaa atacaatgat tacctagaag tgcaagaaat
     3421 gaccgacatc acgtggctca actcccccag tcaggtggcg gtagtgccct gggggttgcg
     3481 caaagtgctg aactggctga agttcaagta cggagacctc cccatgtaca taatatccaa
     3541 tggaatcgat gacgggctgc atgctgagga cgaccagctg agggtgtatt atatgcagaa
     3601 ttacataaac gaagctctca aagcccacat actggatggt atcaatcttt gcggatactt
     3661 tgcttattcg tttaacgacc gcacagctcc gaggtttggc ctctatcgtt atgctgcaga
     3721 tcagtttgag cccaaggcat ccatgaaaca ttacaggaaa attattgaca gcaatggttt
     3781 cccgggccca gaaactctgg aaagattttg tccagaagaa ttcaccgtgt gtactgagtg
     3841 cagttttttt cacacccgaa agtctttact ggctttcata gcttttctat tttttgcttc
     3901 tattatttct ctctccctta tattttacta ctcgaagaaa ggcagaagaa gttacaaagg
     3961 tggaggcgga tcaggaggcg gtggatctgg atccgccacc atgagtaaag gagaagaact
     4021 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
     4081 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat
     4141 ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt tcacttatgg
     4201 tgttcaatgc ttttcaagat acccagatca tatgaaacgg catgactttt tcaagagtgc
     4261 catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa
     4321 gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
     4381 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
     4441 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat
     4501 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc
     4561 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     4621 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     4681 tgggattaca catggcatgg atgaactata caaatagggg cccgtttaaa cccgctgatc
     4741 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4801 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4861 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4921 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4981 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     5041 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5101 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5161 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5221 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5281 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5341 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5401 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5461 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5521 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5581 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5641 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5701 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5761 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5821 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5881 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5941 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6001 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6061 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6121 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6181 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6241 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6301 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6361 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6421 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6481 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6541 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6601 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6661 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6721 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6781 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6841 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6901 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6961 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     7021 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     7081 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     7141 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     7201 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7261 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7321 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7801 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7921 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7981 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8041 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8101 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8161 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8221 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8281 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8341 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8401 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8461 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8521 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8581 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8641 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8701 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8761 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8821 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8881 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8941 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9001 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9061 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9121 catttccccg aaaagtgcca cctgacgtc
//
