LOCUS       V018931                 9073 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018931
VERSION     V018931
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             1678..4560
                     /label="GABBR1(NM_001470)"
                     /note="GABBR1(NM_001470)"
                     /gene="GABBR1"
     polyA_signal    4640..4864
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4910..5338
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5352..5681
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5748..6542
                     /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    6716..6837
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6886..6902)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6910..6926
                     /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(6934..6964)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6979..7000
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7288..7873)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8044..8904)
                     /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(8905..9009)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagcttg gtaccgagct cggatccatg
     1681 ttgctgctgc tgctactggc gccactcttc ctccgccccc cgggcgcggg cggggcgcag
     1741 acccccaacg ccacctcaga aggttgccag atcatacacc cgccctggga agggggcatc
     1801 aggtaccggg gcctgactcg ggaccaggtg aaggctatca acttcctgcc agtggactat
     1861 gagattgagt atgtgtgccg gggggagcgc gaggtggtgg ggcccaaggt ccgcaagtgc
     1921 ctggccaacg gctcctggac agatatggac acacccagcc gctgtgtccg aatctgctcc
     1981 aagtcttatt tgaccctgga aaatgggaag gttttcctga cgggtgggga cctcccagct
     2041 ctggacggag cccgggtgga tttccggtgt gaccccgact tccatctggt gggcagctcc
     2101 cggagcatct gtagtcaggg ccagtggagc acccccaagc cccactgcca ggtgaatcga
     2161 acgccacact cagaacggcg cgcagtgtac atcggggcac tgtttcccat gagcgggggc
     2221 tggccagggg gccaggcctg ccagcccgcg gtggagatgg cgctggagga cgtgaatagc
     2281 cgcagggaca tcctgccgga ctatgagctc aagctcatcc accacgacag caagtgtgat
     2341 ccaggccaag ccaccaagta cctatatgag ctgctctaca acgaccctat caagatcatc
     2401 cttatgcctg gctgcagctc tgtctccacg ctggtggctg aggctgctag gatgtggaac
     2461 ctcattgtgc tttcctatgg ctccagctca ccagccctgt caaaccggca gcgtttcccc
     2521 actttcttcc gaacgcaccc atcagccaca ctccacaacc ctacccgcgt gaaactcttt
     2581 gaaaagtggg gctggaagaa gattgctacc atccagcaga ccactgaggt cttcacttcg
     2641 actctggacg acctggagga acgagtgaag gaggctggaa ttgagattac tttccgccag
     2701 agtttcttct cagatccagc tgtgcccgtc aaaaacctga agcgccagga tgcccgaatc
     2761 atcgtgggac ttttctatga gactgaagcc cggaaagttt tttgtgaggt gtacaaggag
     2821 cgtctctttg ggaagaagta cgtctggttc ctcattgggt ggtatgctga caattggttc
     2881 aagatctacg acccttctat caactgcaca gtggatgaga tgactgaggc ggtggagggc
     2941 cacatcacaa ctgagattgt catgctgaat cctgccaata cccgcagcat ttccaacatg
     3001 acatcccagg aatttgtgga gaaactaacc aagcgactga aaagacaccc tgaggagaca
     3061 ggaggcttcc aggaggcacc gctggcctat gatgccatct gggccttggc actggccctg
     3121 aacaagacat ctggaggagg cggccgttct ggtgtgcgcc tggaggactt caactacaac
     3181 aaccagacca ttaccgacca aatctaccgg gcaatgaact cttcgtcctt tgagggtgtc
     3241 tctggccatg tggtgtttga tgccagcggc tctcggatgg catggacgct tatcgagcag
     3301 cttcagggtg gcagctacaa gaagattggc tactatgaca gcaccaagga tgatctttcc
     3361 tggtccaaaa cagataaatg gattggaggg tcccccccag ctgaccagac cctggtcatc
     3421 aagacattcc gcttcctgtc acagaaactc tttatctccg tctcagttct ctccagcctg
     3481 ggcattgtcc tagctgttgt ctgtctgtcc tttaacatct acaactcaca tgtccgttat
     3541 atccagaact cacagcccaa cctgaacaac ctgactgctg tgggctgctc actggcttta
     3601 gctgctgtct tccccctggg gctcgatggt taccacattg ggaggaacca gtttcctttc
     3661 gtctgccagg cccgcctctg gctcctgggc ctgggcttta gtctgggcta cggttccatg
     3721 ttcaccaaga tttggtgggt ccacacggtc ttcacaaaga aggaagaaaa gaaggagtgg
     3781 aggaagactc tggaaccctg gaagctgtat gccacagtgg gcctgctggt gggcatggat
     3841 gtcctcactc tcgccatctg gcagatcgtg gaccctctgc accggaccat tgagacattt
     3901 gccaaggagg aacctaagga agatattgac gtctctattc tgccccagct ggagcattgc
     3961 agctccagga agatgaatac atggcttggc attttctatg gttacaaggg gctgctgctg
     4021 ctgctgggaa tcttccttgc ttatgagacc aagagtgtgt ccactgagaa gatcaatgat
     4081 caccgggctg tgggcatggc tatctacaat gtggcagtcc tgtgcctcat cactgctcct
     4141 gtcaccatga ttctgtccag ccagcaggat gcagcctttg cctttgcctc tcttgccata
     4201 gttttctcct cctatatcac tcttgttgtg ctctttgtgc ccaagatgcg caggctgatc
     4261 acccgagggg aatggcagtc ggaggcgcag gacaccatga agacagggtc atcgaccaac
     4321 aacaacgagg aggagaagtc ccggctgttg gagaaggaga accgtgaact ggaaaagatc
     4381 attgctgaga aagaggagcg tgtctctgaa ctgcgccatc aactccagtc tcggcagcag
     4441 ctccgctccc ggcgccaccc accgacaccc ccagaaccct ctgggggcct gcccagggga
     4501 ccccctgagc cccccgaccg gcttagctgt gatgggagtc gagtgcattt gctttataag
     4561 tgagaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     4621 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4681 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4741 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4801 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4861 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4981 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5221 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5281 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     5341 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5401 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5461 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5521 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5581 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     5641 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5701 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5761 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5821 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5881 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5941 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6001 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6061 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6121 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6181 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6241 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     6301 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6361 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6421 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6481 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6541 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6601 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6661 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6721 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6781 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6841 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6901 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6961 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7081 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7801 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7861 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7921 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7981 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8881 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9001 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     9061 gatcccctat ggt
//
