LOCUS       V022633                 7284 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022633
VERSION     V022633
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             884..1600
                     /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             1622..2773
                     /label="Gpr88(NM_022427)"
                     /note="Gpr88(NM_022427)"
                     /gene="Gpr88"
     polyA_signal    2851..3075
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3121..3549
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3563..3892
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3959..4753
                     /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    4927..5048
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5097..5113)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5121..5137
                     /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(5145..5175)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5190..5211
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5499..6084)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6255..7115)
                     /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(7116..7220)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      961 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1021 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1081 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1141 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1201 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1261 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1321 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1381 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1441 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1501 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1561 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc
     1621 tatgaccaac tcctcctcca cttcgacctc caccactacc gggggatcgc tgctgctgct
     1681 ctgcgaggaa gaagaatcgt gggcgggccg gcggatcccg gtgtctctcc tgtactcggg
     1741 cctggccatc gggggcacgc tggccaacgg tatggtcatc tatctcgtgt cgtccttccg
     1801 aaagctgcag accaccagca acgccttcat cgtgaatggc tgcgccgcgg atctcagcgt
     1861 ctgcgccctc tggatgccac aggaggcggt gctggggctc ctgccctcgg gctccgcgga
     1921 gcccccgggg gactgggacg gtggcggagg cagctaccgc ctgctccggg gcgggttgtt
     1981 gggcctcggg ctcactgtgt cccttctgtc ccattgcctc gtggcgctga accgctacct
     2041 gctcatcacc agggcgcctg ccacctacca ggtgctctac caacggcgcc acacggtggg
     2101 catgctggca ctgtcctggg cgctcgcctt gggtctggtg ctgctgctcc cgccctgggc
     2161 tcctaagccc ggagccgagc caccgcaagt ccactacccc gcgctcctgg cggcgggagc
     2221 gctgcttgcg cagacggcgc tgctgttgca ctgctacctg ggcatcgtgc gccgcgtgcg
     2281 cgtcagcgtc aagcgggtca gcgtgctcaa cttccacctg ctgcaccagc tacccggctg
     2341 cgccgccgcc gccgccgcct tccccgccgc cccgcacgcg ccgggtccgg gaggcgccgc
     2401 gcatcccgcg cagcctcaac ccctgccggc cgctctgcag cctcgccggg cgcagcggcg
     2461 tctcagcggc ttgtcggtgc tgctgctctg ctgcgtcttc ctgctggcca cgcagccgct
     2521 ggtgtgggtg agcctggcca gtggcttctc cctcccggtg ccctggggcg tgcaggcggc
     2581 cagctggctc ctgtgctgcg ccctgtcggc gctcaacccg ctgctctaca cgtggaggaa
     2641 cgaggagttc cgcagatccg tgcgatctgt cctgccgggc gtcggggacg cggctgcggc
     2701 cgccgcggcc gccaccgcgg tgccagccat gtcccaggcg caactgggta cccgggcggc
     2761 tggccagcac tggtaagaat tctgcagtcg acggtaccgc gggcccggga tcctctagag
     2821 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2881 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2941 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3001 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3061 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3121 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3181 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3241 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3301 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3361 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3421 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3481 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3541 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3601 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3661 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3721 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3781 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3841 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3901 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3961 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4021 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4081 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4141 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4201 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4261 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4321 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4381 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4441 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4501 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4561 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4621 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4681 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4741 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4801 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4861 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4921 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4981 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5041 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5101 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5161 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5221 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5281 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5341 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5401 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5461 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5521 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5581 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5641 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5701 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5761 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5821 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5881 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5941 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6001 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     6061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6121 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6181 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6241 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6301 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6361 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6421 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6481 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6541 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6601 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6661 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6721 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6781 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6841 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6901 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6961 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7021 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7081 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7141 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7201 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7261 ggagatctcc cgatccccta tggt
//
