LOCUS       V032980                 7993 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032980
VERSION     V032980
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..3483
                     /label="Slitrk6(NM_175499)"
                     /note="Slitrk6(NM_175499)"
                     /gene="Slitrk6"
     CDS             3490..3555
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3593..3817
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3863..4291
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4305..4634
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4701..5495
                     /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    5669..5790
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5839..5855)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5863..5879
                     /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(5887..5917)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5932..5953
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6241..6826)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6997..7857)
                     /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(7858..7962)
                     /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 accatgaagc tgtggactta tctcctttat ccatctcttc ttgcttgtct gtctctacag
     1021 tcccagtcac caatgccctc agtcagaggc tcttgcgaca ctctttgtaa ttgtgaggag
     1081 aaagatggca taatgataat aaactgtgaa gaaaaaggaa tcaataaatt atcccaaatc
     1141 agtgtgccac catcacgacc tttccactta agtttattaa ataatggctt gacaatgctt
     1201 cacacgaacg acttttctgg acttaccaat gccctttcaa tacaccttgg atttaacaat
     1261 atagcagaca tagagactgg tgcatttaat ggccttgggc ttcttaagca acttcatatc
     1321 aatcataatt ctttggaaat tcttaaagag gataccttcc atggtttgga gaacctggaa
     1381 ttcttgcagg ctgataataa tttcattacg ataattgaac caagtgcctt cagcaagctc
     1441 aacaggctaa aggtgttgat tttaaatgac aacgctattg agagtcttcc tccaaacata
     1501 tttcgatttg ttcctttaac ccatctagat cttcgtggga accagttgca aacattgcct
     1561 tatgttggct ttttagaaca cattggccgg atattggatc tccaattgga ggacaataag
     1621 tgggcctgca attgtgaatt gttgcagcta aaaaactggc tagaaaatat gcctccacaa
     1681 tctataattg gtgatgtgat atgctacagc cctccacctt tcaaagggag tgtactgagt
     1741 cgcctgaaaa aggaatcttt ttgccctact ccaccagtat acgaagagca tgaggatcca
     1801 tcaggatctt tgctggccat aacatcttca acaagtgaca gccgcctatc aagcaagaat
     1861 acatccattt tgaaacaacc taccaaagca ccaggtttga taccatacct tacaaagcca
     1921 tccactcagc ttccagtacc ttattgtcct atcccttgca attgcaaagt tctctcccct
     1981 tcggggcttc ttatccactg tcaggagcgc aatatagaaa gcttgtcaga tctacagcct
     2041 cctccacaca accctagaaa gctgattctt gcaggcaaca tcattcacac cttaatgaag
     2101 tctgacctca cagactattt cactttggaa atgcttcact tgggaaacaa ccgcattgaa
     2161 gttctggaag aaggatcgtt tatgaatctg acacgactcc agaagcttta tctcaatggc
     2221 aatcatctga ccaaactcaa taaaggcatg ttccttggtc tccacagtct cgagtatttg
     2281 tatcttgagt acaatgcggt aaaggaaata ttaccaggga ccttcaaccc aatgcctaaa
     2341 ctgaaagttc tttatttaaa taacaatctc cttcaagttt taccggcaca tatcttttta
     2401 ggtattcctc taactagagt aaatcttaaa acaaaccaat tcacccattt acctgtaagc
     2461 aacatcttag atgaccttga cttcctgata cagattgacc ttgaggataa cccctgggac
     2521 tgctcttgtg acttggttgg attgcagcaa tggatccaca agcttggcaa gggcaccatg
     2581 actgatgata tcctctgcac ttccccaggg catcttgaca agaaagaatt aaaagccctc
     2641 aacagtgatc ttctttgccc tggtttagtg aataatcctt ccatgccaac ccagactact
     2701 tatgttattg tcacttctcc cacagtggcg gcagatacag ctagtacact cttcagttct
     2761 ctcactgatg ctgtgcccct gtcagttcta atattaggac tcctgattgt gttcataact
     2821 attgtattct gtgccgcggg gatcgtggtt tttgtcctcc accgtaggag aagatataag
     2881 aagaaaaaag tagaggaaca actgagagac aacagtcctg tacacttgca gtacagcatg
     2941 tatggtcata aaacaaccca ccatacaacg gaaagaccct cttcctctct ctatgaacaa
     3001 cacatggtga gcccaatggt tcatgtctac agaagtccat cctttggtcc aaagcatttg
     3061 gaagaagttg aggaaagaaa tgataaagaa ggaaatgatg caaaacatct tcaaaggagc
     3121 ctgttagaac gagaaaatca ttcaccactc acaggttcaa atatgaaata caaaactaca
     3181 gaccaatcaa cagatttcat atcctttcag gatgcaagct tattatatag gaacattttg
     3241 gagaaagaga gggaacttca gcaacttggc atcacagagt acctaagaaa aaatcttgct
     3301 cagctccagc cagaggtgga ggttaactat cctggagccc acgaggagct aaaattaatg
     3361 gaaacgttga tgtactcacg accaaggaag gtgttggtag aacagactaa aaatgaatac
     3421 tttgagctca aagctaactt gcatgctgaa cccgactact tagaagtctt ggagcagcaa
     3481 acactcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     3541 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3601 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3661 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3721 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3781 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3841 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3901 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3961 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4021 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4081 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4141 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4201 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4261 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4321 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4381 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4441 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4501 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4561 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4621 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4681 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4741 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4801 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4861 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4921 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4981 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5041 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5101 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5161 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5221 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5281 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5341 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5401 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5461 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5521 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5581 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5641 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5701 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5761 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5821 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5881 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5941 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6001 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6061 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6241 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6661 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6901 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6961 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7021 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7081 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7141 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7201 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7261 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7321 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7381 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7441 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7501 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7561 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7621 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7681 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7741 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7801 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7861 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7921 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7981 gccacctgac gtc
//
