LOCUS       V036146                10165 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036146
VERSION     V036146
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..5655
                     /label="PFAS(NM_012393)"
                     /note="PFAS(NM_012393)"
                     /gene="PFAS"
     polyA_signal    5732..5956
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6002..6430
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6444..6773
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6840..7634
                     /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    7808..7929
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7978..7994)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8002..8018
                     /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(8026..8056)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8071..8092
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8380..8965)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9136..9996)
                     /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(9997..10101)
                     /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 catgtcccca gtccttcact tctatgttcg tccctctggc
     1681 catgaggggg cagcccctgg acacactcgg aggaaactgc aagggaaact gccagagctg
     1741 cagggcgtcg agactgaact gtgctacaac gtgaactgga cagctgaggc cctccccagt
     1801 gctgaggaga caaagaagct gatgtggctg tttggttgcc ccttactgct ggatgatgtt
     1861 gctcgggagt cctggctcct tcctggctcc aatgacctgc tgctggaggt cgggcccagg
     1921 ctgaacttct ccaccccaac atccaccaac atcgtgtcag tgtgccgcgc cactgggctg
     1981 gggcctgtgg atcgtgtgga gaccacccgg cgctaccggc tctcgtttgc ccaccccccg
     2041 tcagctgagg tggaagccat tgctctggct accctgcacg accggatgac agagcagcac
     2101 ttcccccatc ccatccagag tttctcccct gagagcatgc cggaacccct caatggccct
     2161 atcaatatac tgggtgaggg ccggcttgcg ctggagaagg ccaaccagga gcttggtctg
     2221 gctttagact cttgggacct agacttctac accaagcgct tccaggagct acagcggaac
     2281 ccgagcactg tggaggcctt tgacttggcg cagtccaata gcgagcacag ccgacactgg
     2341 ttcttcaagg gccagctcca cgtggatggg cagaagctgg tgcactcact gtttgagtcc
     2401 atcatgagca cccaggaatc ctcgaacccc aacaacgtcc tcaaattctg tgataacagc
     2461 agtgcaatcc agggaaagga agtccgattc ctacggcctg aggaccccac acggccaagc
     2521 cgcttccagc aacagcaagg gctgagacat gttgtcttca cagcagagac tcacaacttt
     2581 cccacaggag tatgcccctt tagtggtgca accactggca cagggggccg gattcgagat
     2641 gtccagtgca caggccgcgg ggcccacgtg gtggctggca ctgccggcta ttgctttgga
     2701 aatctgcata ttccaggtta caatctgccc tgggaggatc caagcttcca gtatcctggg
     2761 aattttgccc ggcccctgga ggttgccatt gaagccagta atggagcttc tgactatggc
     2821 aacaagtttg gggaaccagt gctggctggc ttcgcccgct ccttgggcct ccagctccca
     2881 gacggccagc ggcgtgagtg gatcaagccc atcatgttta gtgggggcat tgggtccatg
     2941 gaagctgacc acataagcaa ggaggcccca gagccaggca tggaagttgt aaaggttgga
     3001 ggtcccgtct acaggattgg agttggaggt ggagctgctt catctgtgca ggtgcaggga
     3061 gataacacca gtgacctgga ctttggggct gtgcagcgag gagacccgga gatggaacag
     3121 aagatgaacc gtgtgatcag ggcttgtgtg gaggccccca agggaaaccc catctgcagc
     3181 cttcatgatc agggcgctgg tggcaatggc aatgtcctaa aagagctgag tgacccagct
     3241 ggagccatca tttacaccag ccgcttccag cttggggacc caaccctgaa tgccctggaa
     3301 atctgggggg ctgagtacca ggaatcaaat gctcttctgc tgaggtcccc caaccgggac
     3361 ttcctgactc atgtcagtgc ccgtgaacgt tgcccggctt gcttcgtggg caccatcact
     3421 ggagaccgga gaatagtgct ggtggacgat cgggagtgtc ctgtcagaag aaatggccag
     3481 ggggatgccc ccccgacacc cctgccaacc cctgtggacc tggagctcga atgggtgctg
     3541 ggcaagatgc ctcggaagga gttcttcctg cagaggaagc cccccatgct gcagcctctg
     3601 gccttgcccc cagggctgag cgtgcaccag gctctggaga gggttctgag gctgcccgcc
     3661 gtggccagca agcgctacct caccaataag gtggaccgct ctgtgggagg cctggtggcc
     3721 cagcagcagt gcgtggggcc cctgcaaact cctctggcag atgtagcggt tgtggcactg
     3781 agccatgagg agctcatagg ggctgccaca gccttgggag aacagccagt caagagcctg
     3841 ctggacccaa aagtcgccgc ccggctggcc gtggccgaag ccctcaccaa cctggtgttt
     3901 gctctggtca ctgacctccg ggatgtgaag tgtagcggga actggatgtg ggcagccaag
     3961 ctcccagggg agggcgcagc tttggcggat gcctgtgagg ctatggtggc agtgatggca
     4021 gccctgggtg tggcagtgga tggtggcaag gactccctca gcatggctgc tcgggttggc
     4081 actgagaccg tgcgggctcc tgggtcactg gtcatctcag cctatgccgt ctgcccagac
     4141 atcacagcca ctgtgacccc agacctcaag catcctgaag ggagaggcca tctgctctat
     4201 gtggctctga gccctgggca gcaccggctc gggggcacag ctctggccca gtgcttctcc
     4261 cagcttgggg aacaccctcc agacctggac cttcctgaga acttggtgcg ggccttcagc
     4321 atcactcagg ggctgctgaa agaccgcctc ctctgctcag gccacgatgt cagtgacgga
     4381 ggcctcgtca catgcctgct ggagatggcc tttgctggaa attgcgggct acaggtggat
     4441 gtgcctgtcc ccagggttga tgtcctgtct gtgctgttcg ctgaggagcc aggcctcgtg
     4501 ctggaggtgc aggagccaga cctggcccag gtgctgaagc gttaccggga tgctggcctc
     4561 cattgcctgg agctgggcca cacaggcgag gccgggcccc acgccatggt ccgggtgtca
     4621 gtgaacgggg ctgtggttct ggaggagcct gttggggagc tgcgagccct ctgggaggag
     4681 acgagtttcc agctggaccg gctacaggca gagcctcgct gtgtggcaga ggaggaacgg
     4741 ggcctgaggg agcggatggg gcccagctat tgcctgcccc ccacctttcc caaagcctcc
     4801 gtgccccgtg agcctggtgg tcccagcccc cgagtcgcca tcttgcgaga ggagggcagt
     4861 aatggagacc gggagatggc cgatgccttc cacttagctg ggtttgaggt atgggacgtg
     4921 accatgcagg acctctgctc tggggcaatt gggctggaca ctttccgtgg cgtggccttc
     4981 gtgggcggct tcagctatgc agatgtcctg ggctctgcca aagggtgggc agctgctgtg
     5041 acctttcatc ccagggctgg ggctgagctg aggcgcttcc ggaagcggcc agacaccttc
     5101 agcctgggcg tgtgtaatgg ctgtcaactg ctggctctgc tcggctgggt gggaggcgac
     5161 cccaatgagg atgctgcaga gatgggccct gactcccagc cagcccggcc aggccttctg
     5221 ctacgccaca acctgtctgg gcgctacgag tctcgctggg ccagcgtgcg tgtggggcct
     5281 gggccagccc tgatgctgcg agggatggag ggcgccgtgc tgcccgtgtg gagtgcgcac
     5341 ggggaaggtt acgtagcatt ttcttctccg gaactccaag ctcagattga ggccaggggc
     5401 ttggctccac tgcactgggc tgatgatgac gggaacccca cagagcagta ccctctgaat
     5461 cccaatgggt ccccaggggg cgtggctggc atctgctcct gtgatggccg ccacctggct
     5521 gtcatgcctc accctgagcg ggccgttagg ccttggcagt gggcatggcg accccctcca
     5581 tttgatactc tgaccacctc cccctggctc cagctcttta tcaatgcccg aaactggacc
     5641 ctggaaggga gctgctgagg atcctgatat ccagcacagt ggcggccgct cgagtctaga
     5701 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     5761 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     5821 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     5881 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     5941 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     6001 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6061 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6421 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     6481 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6601 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6661 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     6721 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     6781 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     6841 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6901 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6961 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     7021 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     7081 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     7141 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     7201 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     7261 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     7321 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     7381 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     7441 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     7501 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7561 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7621 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     7681 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7741 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     7801 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     7861 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     7921 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     7981 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     8041 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     8101 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     8161 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     8221 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     8281 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8341 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8401 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     8461 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     8521 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     8581 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     8641 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     8701 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     8761 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     8821 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     8881 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     8941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     9001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     9061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     9121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     9181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     9241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    10021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    10081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    10141 gggagatctc ccgatcccct atggt
//
