LOCUS       V030222                 7442 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030222
VERSION     V030222
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             956..982
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             983..1006
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1007..2962
                     /label="ADGRE3(NM_032571)"
                     /note="ADGRE3(NM_032571)"
                     /gene="ADGRE3"
     polyA_signal    3009..3233
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3279..3707
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3721..4050
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4117..4911
                     /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    5085..5206
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5255..5271)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5279..5295
                     /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(5303..5333)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5348..5369
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5657..6242)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6413..7273)
                     /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(7274..7378)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg aagaccatca tcgccctgag ctacatcttc tgcctggtgt tcgcctaccc
      961 atacgacgtc ccagactacg ctgactacaa ggacgatgat gacaaaatgc agggaccatt
     1021 gcttcttcca ggcctctgct ttctgctgag cctctttgga gctgtgactc agaaaaccaa
     1081 aacttcctgt gctaagtgcc ccccaaatgc ttcctgtgtc aataacactc actgcacctg
     1141 caaccatgga tatacttctg gatctgggca gaaactattc acattcccct tggagacatg
     1201 taacgacatt aatgaatgta caccacccta tagtgtatat tgtggattta acgctgtgtg
     1261 ttacaatgtc gaaggaagtt tctactgtca atgtgtccca ggatatagac tgcattctgg
     1321 gaatgaacaa ttcagtaatt ccaatgagaa cacctgtcag gacaccacct cctcaaagac
     1381 aaccgagggc aggaaagagc tgcaaaagat tgtggacaaa tttgagtcac ttctcaccaa
     1441 tcagacttta tggagaacag aagggagaca agaaatctca tccacagcta ccactattct
     1501 ccgggatgtg gaatcgaaag ttctagaaac tgccttgaaa gatccagaac aaaaagtcct
     1561 gaaaatccaa aacgatagtg tagctattga aactcaagcg attacagaca attgctctga
     1621 agaaagaaag acattcaact tgaacgtcca aatgaactca atggacatcc gttgcagtga
     1681 catcatccag ggagacacac aaggtcccag tgccattgcc tttatctcat attcttctct
     1741 tggaaacatc ataaatgcaa ctttttttga agagatggat aagaaagatc aagtgtatct
     1801 gaactctcag gttgtgagtg ctgctattgg acccaaaagg aacgtgtctc tctccaagtc
     1861 tgtgacgctg actttccagc acgtgaagat gacccccagt accaaaaagg tcttctgtgt
     1921 ctactggaag agcacagggc agggcagcca gtggtccagg gatggctgct tcctgataca
     1981 cgtgaacaag agtcacacca tgtgtaattg cagtcacctg tccagcttcg ctgtcctgat
     2041 ggccctgacc agccaggagg aggatcccgt gctgactgtc atcacctacg tggggctgag
     2101 cgtctctctg ctgtgcctcc tcctggcggc cctcactttt ctcctgtgta aagccatccg
     2161 gaacaccagc acctcactgc atctgcagct ctcgctctgc ctcttcctgg cccacctcct
     2221 cttcctcgtg gggattgatc gaactgaacc caaggtgctg tgctccatca tcgccggtgc
     2281 tttgcactat ctctacctgg ccgccttcac ctggatgctg ctggagggtg tgcacctctt
     2341 cctcactgca cggaacctga cagtggtcaa ctactcaagc atcaatagac tcatgaagtg
     2401 gatcatgttc ccagtcggct atggcgttcc cgctgtgact gtggccattt ctgcagcctc
     2461 ctggcctcac ctttatggaa ctgctgatcg atgctggctc cacctggacc agggattcat
     2521 gtggagtttc cttggcccag tctgtgccat tttctctgcg aatttagtat tgtttatctt
     2581 ggtcttttgg attttgaaaa gaaaactttc ctccctcaat agtgaagtgt caaccatcca
     2641 gaacacaagg atgctggctt tcaaagcaac agctcagctc ttcatcctgg gctgcacatg
     2701 gtgtctgggc ttgctacagg tgggtccagc tgcccaggtc atggcctacc tcttcaccat
     2761 catcaacagc ctccaaggct tcttcatctt cttggtctac tgcctcctca gccagcaggt
     2821 ccagaaacaa tatcaaaagt ggtttagaga gatcgtaaaa tcaaaatctg agtctgagac
     2881 atacacactt tccagcaaga tgggtcctga ctcaaaaccc agtgaggggg atgtttttcc
     2941 aggacaagtg aagagaaaat attaactcga gtctagaggg cccgtttaaa cccgctgatc
     3001 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3061 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3121 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3181 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3241 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3301 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3361 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3421 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3481 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3541 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3601 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3661 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3721 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3781 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3841 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3901 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3961 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4021 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4081 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4141 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4201 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4261 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4321 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4381 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4441 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4501 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4561 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4621 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4681 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4741 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4801 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4861 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4921 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4981 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5041 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5101 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5161 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5221 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5281 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5341 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5401 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5461 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5521 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5581 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5641 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5701 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5761 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5821 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5881 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5941 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6001 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6061 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6121 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6181 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6361 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6421 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6481 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6541 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6601 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6661 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6721 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6781 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6841 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6901 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6961 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7021 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7081 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7141 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7201 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7261 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7321 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7381 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7441 gt
//
