LOCUS       V038256                10009 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038256
VERSION     V038256
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..4668
                     /label="ADGRA3(NM_145290)"
                     /note="ADGRA3(NM_145290)"
                     /gene="ADGRA3"
     CDS             4675..4701
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4705..4731
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4735..4761
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    4785..5373
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5439..5487
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5689..6117
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6131..6460
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6527..7321
                     /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    7497..7618
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7667..7683)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7691..7707
                     /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(7715..7745)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7760..7781
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8069..8657)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8828..9688)
                     /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(9689..9793)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga gccacccgga
      721 cgccggcggg gccgcgcgca gccgccgctg ttgctgccgc tctcgctgtt agcgctgctc
      781 gcgctgctgg gaggcggcgg cggcggcggc gccgcggcgc tgcccgccgg ctgcaagcac
      841 gatgggcggc cccgaggggc tggcagggcg gcgggcgccg ccgagggcaa ggtggtgtgc
      901 agcagcctgg aactcgcgca ggtcctgccc ccagatactc tgcccaaccg cacggtcacc
      961 ctgattctga gtaacaataa gatatccgag ctgaagaatg gctcattttc tgggttaagt
     1021 ctccttgaaa gattggacct ccgaaacaat cttattagta gtatagatcc aggtgccttc
     1081 tggggactgt catctctaaa aagattggat ctgacaaaca atcgaatagg atgtctgaat
     1141 gcagacatat ttcgaggact caccaatctg gttcggctaa acctttcggg gaatttgttt
     1201 tcttcattat ctcaaggaac ttttgattat cttgcgtcat tacggtcttt ggaattccag
     1261 actgagtatc ttttgtgtga ctgtaacata ctgtggatgc atcgctgggt aaaggagaag
     1321 aacatcacgg tacgggatac caggtgtgtt tatcctaagt cactgcaggc ccaaccagtc
     1381 acaggcgtga agcaggagct gttgacatgc gaccctccgc ttgaattgcc gtctttctac
     1441 atgactccat ctcatcgcca agttgtgttt gaaggagaca gccttccttt ccagtgcatg
     1501 gcttcatata ttgatcagga catgcaagtg ttgtggtatc aggatgggag aatagttgaa
     1561 accgatgaat cgcaaggtat ttttgttgaa aagaacatga ttcacaactg ctccttgatt
     1621 gcaagtgccc taaccatttc taatattcag gctggatcta ctggaaattg gggctgtcat
     1681 gtccagacca aacgtgggaa taatacgagg actgtggata ttgtggtatt agagagttct
     1741 gcacagtact gtcctccaga gagggtggta aacaacaaag gtgacttcag atggcccaga
     1801 acattggcag gcattactgc atatctgcag tgtacgcgga acacccatgg cagtgggata
     1861 tatcccggaa acccacagga tgagagaaaa gcttggcgca gatgtgatag aggtggcttt
     1921 tgggcagatg atgattattc tcgctgtcag tatgcaaatg atgtcactag agttctttat
     1981 atgtttaatc agatgcccct caatcttacc aatgccgtgg caacagctcg acagttactg
     2041 gcttacactg tggaagcagc caacttttct gacaaaatgg atgttatatt tgtggcagaa
     2101 atgattgaaa aatttggaag atttaccaag gaggaaaaat caaaagagct aggtgacgtg
     2161 atggttgaca ttgcaagtaa catcatgttg gctgatgaac gtgtcctgtg gctggcgcag
     2221 agggaagcta aagcctgcag taggattgtg cagtgtcttc agcgcattgc tacctaccgg
     2281 ctagccggtg gagctcacgt ttattcaaca tattcaccca atattgctct ggaagcttat
     2341 gtcatcaagt ctactggctt cacggggatg acctgtaccg tgttccagaa agtggcagcc
     2401 tctgatcgta caggactttc ggattatggg aggcgggatc cagagggaaa cctggataag
     2461 cagctgagct ttaagtgcaa tgtttcaaat acattttcga gtctggcact aaagaatact
     2521 attgtggagg cttctattca gcttcctcct tcccttttct caccaaagca aaaaagagaa
     2581 ctcagaccaa ctgatgactc tctttacaag cttcaactca ttgcattccg caatggaaag
     2641 ctttttccag ccactggaaa ttcaacaaat ttggctgatg atggaaaacg acgtactgtg
     2701 gttacccctg tgattctcac caaaatagat ggtgtgaatg tagataccca ccacatccct
     2761 gttaatgtga cactgcgtcg aattgcacat ggagcagatg ctgttgcagc ccggtgggat
     2821 ttcgatttgc tgaacggaca aggaggctgg aagtcagatg ggtgccatat actctattca
     2881 gatgaaaata tcactacgat tcagtgctac tcccttagta actatgcagt tttaatggat
     2941 ttgacgggat ctgaactata cacccaggcg gccagcctcc tgcatcctgt ggtttatact
     3001 accgctatca ttctcctctt atgtctctta gccgtcattg tcagttacat ataccatcac
     3061 agtttgatta gaatcagcct caagagctgg cacatgcttg tgaacttgtg ctttcatatt
     3121 ttcctaacct gtgtggtctt tgtgggagga ataacccaga ctaggaatgc cagcatctgc
     3181 caagcagttg ggataattct tcactattcc acccttgcca cagtactatg ggtaggagtg
     3241 acagctcgaa atatctacaa acaagtcact aaaaaagcta aaagatgcca ggatcctgat
     3301 gaaccaccac ctccaccaag accaatgctc agattttacc tgattggtgg tggtatcccc
     3361 atcattgttt gcggcataac tgcagcagcg aacattaaga attacggcag tcggccaaac
     3421 gcaccctatt gctggatggc atgggaaccc tccttgggag ccttctatgg gccagccagc
     3481 ttcatcactt ttgtaaactg catgtacttt ctgagcatat ttattcagtt gaaaagacac
     3541 cctgagcgca aatatgagct taaggagccc acggaggagc aacagagatt ggcagccaat
     3601 gaaaatggcg aaataaatca tcaggattca atgtctttgt ctctgatttc tacatcagcc
     3661 ttggaaaatg agcacacttt tcattctcag ctcttggggg ccagccttac tttgctctta
     3721 tatgttgcac tgtggatgtt tggggctttg gctgtttctt tgtattaccc tttggacttg
     3781 gtttttagct tcgtttttgg agccacaagt ttaagcttca gtgcgttctt cgtggtccac
     3841 cattgtgtta atagggagga tgttagactt gcgtggatca tgacttgctg cccaggacgg
     3901 agctcgtatt cagtgcaagt caacgtccag ccccccaact ctaatgggac gaatggagag
     3961 gcacccaaat gccccaatag cagtgcggag tcttcatgca caaacaaaag tgcttcaagc
     4021 ttcaaaaatt cctcccaggg ctgcaaatta acaaacttgc aggcggctgc agctcagtgc
     4081 catgccaatt ctttaccttt gaactccacc cctcagcttg ataatagtct gacagaacat
     4141 tcaatggaca atgatattaa aatgcacgtg gcgcctttag aagttcagtt tcgaacaaat
     4201 gtgcactcaa gccgccacca taaaaacaga agtaaaggac accgggcaag ccgactcaca
     4261 gtcctgagag aatatgccta cgatgtccca acgagcgtgg aaggaagcgt gcagaacggc
     4321 ttacctaaaa gccggctggg caataacgaa ggacactcga ggagccgaag agcttattta
     4381 gcctacagag agagacagta caacccaccc cagcaagaca gcagcgatgc ttgtagcaca
     4441 cttcccaaaa gtagcagaaa ttttgaaaag ccagtttcaa ccactagtaa aaaagatgcg
     4501 ttaaggaagc cagctgtggt tgaacttgaa aatcagcaaa aatcttatgg cctcaacttg
     4561 gccattcaga atggaccaat taaaagcaat gggcaggagg gacccttgct cggtaccgat
     4621 agcactggca atgttaggac tggattatgg aaacacgaaa ctactgtgct cgagtaccca
     4681 tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc tagctatcct
     4741 tatgatgtcc cagactacgc atagtgaggg cccgatatct cgacaatcaa cctctggatt
     4801 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4861 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4921 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4981 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     5041 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     5101 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     5161 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     5221 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     5281 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     5341 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac
     5401 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5461 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5521 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5581 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5641 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     5701 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5761 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5821 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc
     5881 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5941 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6001 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg
     6061 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     6121 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6181 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6361 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     6421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     6481 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     6541 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     6601 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     6661 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     6721 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     6781 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     6841 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     6901 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     6961 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     7021 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     7081 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     7141 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     7201 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     7261 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     7321 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     7381 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     7441 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     7501 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7561 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7621 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     7681 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7741 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7801 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7861 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7921 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7981 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     8041 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     8101 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     8161 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     8221 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     8281 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     8341 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     8401 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     8461 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     8521 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     8581 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     8641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     8701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     9001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     9061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     9121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     9181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     9241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     9301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     9361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     9421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     9481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     9541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     9601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     9661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     9721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     9781 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     9841 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
     9901 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     9961 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
