LOCUS       V032218                 6628 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032218
VERSION     V032218
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             964..2118
                     /label="Gpr137b(NM_031999)"
                     /note="Gpr137b(NM_031999)"
                     /gene="Gpr137b"
     CDS             2125..2190
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2228..2452
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2498..2926
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2940..3269
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3336..4130
                     /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    4304..4425
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4474..4490)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4498..4514
                     /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(4522..4552)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4567..4588
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4876..5461)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5632..6492)
                     /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(6493..6597)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggagg cgcctccgtg ggagccggtg cgcaatgact ccctgcctcc cacgctgagc
     1021 cccgcggtgc cgccctacgt gaagctcggc ctcaccgcgg tctacaccgt cttctacgcg
     1081 ctcctcttcg tgttcatcta tgcgcagctc tggctggtgc tgcgctaccg tcacaagcgg
     1141 ctcagctacc agagcgtctt cctcttcctt tgcctcttct gggcctcgct gcgtaccgtg
     1201 ctcttctcct tttacttccg agacttcgtg gcagccaact cgttcagccc cttcgtcttc
     1261 tggctgctct actgcttccc cgtgtgtcta cagttcttca ccctcacgct catgaacttg
     1321 tacttcacgc aggtgatttt caaggccaag tcaaaatatt ctccagagct actcaaatac
     1381 cggttacccc tctacctggc ctcacttttc atcagcctcg ttttcctgtt ggtgaatctg
     1441 acctgtgctg tgctggtgaa gacgggagac tgggacagga aggttatcgt ctctgtgaga
     1501 gtggccatca atgacacact ctttgtgctg tgtgctatct ctctctccat ctgcctctac
     1561 aaaatctcca agatgtccct ggccaacatc tacttggagt caaagggctc atcagtgtgt
     1621 caggtaactg ccattggtgt caccgtcatc ttgctctaca cctctcgggc ctgctacaac
     1681 ctgttcatct tgtcattttc tcagatcaag aacgtccatt cctttgatta tgactggtac
     1741 aatgtatccg accaggcaga tctgaagagc cagctgggtg acgccggcta cgtagtgttt
     1801 ggcgtggtgc ttttcgtgtg ggagctccta cccaccacct tggtggttta tttcttccga
     1861 gtcagaaatc ccacgaagga tcttaccaat cctgggatgg tccccagcca tggattcagt
     1921 cccagatctt acttctttga caacccccga agatatgaca gtgatgatga ccttgcctgg
     1981 aacattgccc ctcagggact tcagggaagt tttgctccag actactatga ttggggacaa
     2041 caaaataaca gcttcctggc acaagcagga actttgcatc aagactccac tttggatcca
     2101 gacaaagcaa gccaagggct cgaggactac aaagaccatg acggtgatta taaagatcat
     2161 gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac ccgctgatca
     2221 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2281 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2341 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2401 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2461 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2521 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2581 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2641 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2701 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2761 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2821 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2881 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2941 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3001 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3061 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3121 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3181 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3241 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3301 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3361 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3421 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3481 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3541 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3601 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3661 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3721 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3781 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3841 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3901 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3961 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4021 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4081 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4141 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4201 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4261 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4321 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4381 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4441 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4501 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4561 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4621 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5401 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5461 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5521 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5581 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5641 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5701 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5761 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5821 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5881 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5941 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6001 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6061 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6121 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6181 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6241 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6301 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6361 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6421 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6481 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6541 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6601 atttccccga aaagtgccac ctgacgtc
//
