LOCUS       V030825                 6545 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030825
VERSION     V030825
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             941..2092
                     /label="Grpr(NM_008177)"
                     /note="Grpr(NM_008177)"
                     /gene="Grpr"
     CDS             2093..2116
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2142..2366
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2412..2840
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2854..3183
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3250..4044
                     /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    4218..4339
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4388..4404)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4412..4428
                     /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(4436..4466)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4481..4502
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4790..5378)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5549..6409)
                     /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(6410..6514)
                     /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 atccgccacc atggctccaa ataattgttc
      961 ccacctgaac ttggacgtgg accctttcct gtcctgcaac gacaccttca atcaaagtct
     1021 gagtcccccc aagatggaca actggtttca cccgggcttc atctatgtca tccctgcagt
     1081 ttatgggctt atcatcgtga taggtcttat tggcaacatc acgctcatca agatcttctg
     1141 cacggtcaag tccatgcgaa acgtgccaaa cctgttcatc tctagcctgg ctttgggaga
     1201 cctgctgctg ctggtgacat gcgcccctgt ggatgccagc aagtacctgg ctgacaggtg
     1261 gctatttggc agaattggct gcaaactgat cccctttata caacttactt cagtgggggt
     1321 gtctgtcttc acacttacgg cactgtcagc tgacaggtac aaagccattg tacggccaat
     1381 ggatatccag gcatcccatg ccctgatgaa gatctgtctc aaagctgctt tgatctggat
     1441 tgtctctatg ttgttggcca tcccagaggc tgtgttttct gacctccacc ccttccatgt
     1501 gaaagatacc aaccaaacct tcattagttg tgccccctac ccacactcca atgagctaca
     1561 ccctaaaatc cattccatgg cttcctttct ggttttctac gttatcccac tggcgatcat
     1621 ctctgtctac tactacttca ttgcccgaaa tctgattcag agtgcctaca atcttcccgt
     1681 ggaaggcaat atacatgtca agaagcagat cgaatcccgg aagcggcttg ccaagacagt
     1741 actggtgttt gtgggcctct ttgccttctg ctggctcccc aaccatgtca tctacctgta
     1801 ccgttcctac cactactctg aagtggacac ctccatgctc cactttgtca ccagcatctg
     1861 tgcccgcctc ctggccttca ccaactcctg tgtgaacccc tttgctcttt atctgctgag
     1921 caagagcttc aggaagcagt tcaacactca acttctctgc tgccagcctg gcctgatgaa
     1981 caggtcccac agcacaggca gaagtaccac ctgcatgacc tccttcaaga gcactaaccc
     2041 ctcggctacc tttagcctca tcaacagaaa tatctgtcat gaggggtatg tcgattacaa
     2101 ggatgacgac gataagtgat aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2161 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2221 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2281 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2341 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2401 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2461 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2521 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2581 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2641 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2701 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2761 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2821 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3001 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3061 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3121 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3181 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3241 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3301 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3361 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3421 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3481 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3541 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3601 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3661 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3721 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3781 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3841 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3901 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3961 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4021 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4081 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4141 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4201 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4321 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4381 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4441 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4501 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4561 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4621 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4681 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4741 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4801 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4861 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4921 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4981 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5041 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5101 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5161 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5221 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5281 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5341 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5401 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5461 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5521 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5581 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5641 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5701 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5761 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5821 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5881 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5941 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6001 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6061 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6121 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6181 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6241 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6301 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6361 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6421 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6481 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6541 acgtc
//
