LOCUS       V018761                10793 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018761
VERSION     V018761
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..6283
                     /label="SIPA1L3(NM_015073)"
                     /note="SIPA1L3(NM_015073)"
                     /gene="SIPA1L3"
     CDS             6290..6355
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    6393..6617
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6663..7091
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7105..7434
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7501..8295
                     /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    8469..8590
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8639..8655)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8663..8679
                     /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(8687..8717)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8732..8753
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(9041..9626)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9797..10657)
                     /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(10658..10762)
                     /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 atgaccacct atcgggccat
      961 ccccagcgat ggtgtggacc tggcagccag ctgtggcgcc agggtgggcg atgtcctccc
     1021 tgggccacac acaggggact acgctccctt gggattctgg gcccagaatg gcagcatgtc
     1081 ccagcctctt ggcgagagcc cggccaccgc caccgccacc gccaccgcca ccacccgccc
     1141 cagccccacc actcccgcaa tgcccaagat gggcgtgcgc gcaagggtgg ccgactggcc
     1201 gcccaagcgg gaggccctga gagagcacag caacccaagc ccctcccagg acacagatgg
     1261 cacaaaggcc accaagatgg cccattccat gaggagcata cagaacggac agccccccac
     1321 cagcaccccg gcttcctcag ggtccaaagc cttccaccga ctctccagga gaaggtccaa
     1381 agacgtggag ttccaggacg ggtggccccg gtcccccggc agggccttcc tcccccttcg
     1441 gcaccgcagc agcagcgaga tcaccctcag cgagtgtgac gcggaggacg cgggggagcc
     1501 gcggggggcc cggcacacgg gggcgctgcc cctcttccgc gagtacggga gcacctcgtc
     1561 catcgacgtg cagggcatgc ccgagcagag cttcttcgac atcctgaacg agttccgcag
     1621 cgagcagccc gacgcccgag ggtgccaggc cctcaccgag ctcctccggg cagatcctgg
     1681 cccacacctc atggggggcg gcggcggagc caagggggac tcccacaacg ggcagcccgc
     1741 caaggacagc ctcctgccac tgcagcccac gaaggagaag gagaaggccc ggaagaaacc
     1801 tgcgcggggc ctcggcggcg gggacacggt ggactcgtcc atctttcgga agctaaggag
     1861 cagcaaaccc gagggggagg ctgggcgttc cccgggggag gccgacgagg gccggagccc
     1921 cccggaagcc agcaggccgt gggtgtgtca gaagagcttc gcccacttcg acgtgcagag
     1981 catgctgttc gacctcaacg aggcggccgc caacagggtg tcggtgtcgc agcggcggaa
     2041 caccaccacg ggtgcttcgg ccgcttccgc cgcctcggcc atggcctccc tcacggcctc
     2101 gcgggcccac agcctcggag gcctggaccc ggccttcacc agcacagagg acctaaactg
     2161 caaggagaac ttggagcagg acctcggcga tgacaacagc aacgacctgc tgctcagctg
     2221 cccgcacttc cgcaatgaga tcgggggcga gtgtgagcgc aacgtgagct tctcccgggc
     2281 ttccgtgggc tccccgagca gcggcgaggg ccacctggca gagcccgccc tgagcgccta
     2341 ccgcaccaac gccagcatct cggtgttgga agttcccaag gagcagcagc ggacgcagag
     2401 tcggccccgg cagtacagca tcgagcatgt ggacctgggc gcccgctact accaggatta
     2461 cttcgtgggc aaagaacatg ccaattactt cggcgtggat gagaagctgg ggccagtggc
     2521 tgtgagcatt aagcgggaga agctggaaga ccacaaggag cacggacctc agtaccagta
     2581 caggatcatc ttccggaccc gcgagctcat caccctgcgg ggctccatcc tggaagatgc
     2641 tacgcccaca gccaccaagc atgggaccgg gcggggcctg cccttgaagg atgccctgga
     2701 gtatgtcatc cccgagctca acatccactg cctgcggctg gccctcaaca cccccaaggt
     2761 gacggagcaa ctgctgaagc tcgatgagca agggctctgc cggaagcaca aggtgggcat
     2821 cctctattgc aaggccggcc agagctccga ggaggagatg tacaacaatg aggaggccgg
     2881 ccccgccttt gaggagttcc tctccctcat cggcgagaag gtctgcctga agggcttcac
     2941 caagtacgct gcccagctgg acgtcaagac cgactccacg ggaacccact ccctctacac
     3001 gatgtaccag gactacgaga tcatgttcca tgtctccacc ctgctccctt acacccccaa
     3061 caacaggcag cagctgctac ggaagaggca cataggaaat gacatcgtga cgatcatctt
     3121 ccaggagcct ggcgcgctac cgttcacccc caagaacatc cgctcccact tccagcacgt
     3181 cttcatcatt gtccgagtcc acaacccctg cactgataac gtctgttaca gtatggctgt
     3241 gacccgatcc aaagacgctc ctcctttcgg cccccccatc cccagtggaa ccacattccg
     3301 caaatccgac gtcttcagag acttcttgct ggccaaggtg attaacgctg agaacgccgc
     3361 gcacaagtcc gacaagttcc acaccatggc caccaggacc cgccaggagt atctcaagga
     3421 cctggccgaa aactgtgtct ccaacacccc catcgactcc accggcaaat tcaacctcat
     3481 ctccctgacc tccaagaaga aggaaaagac aaaagcacgg gctggcgctg agcagcacag
     3541 tgcaggggcc atcgcctgga gggtggtggc ccaggactac gcccaggggg tggaaatcga
     3601 ctgcattttg ggaatttcca atgagtttgt ggtgctcctg gacttacgca ccaaggaggt
     3661 ggtgttcaac tgctactgcg gggatgtcat tggctggact ccagactcct ccacactcaa
     3721 aatcttctat ggacgaggag accacatctt cctacaggcg acagagggtt ctgtggagga
     3781 cataagggag atagtgcaga gactgaaggt gatgaccagt ggctgggaga cggtggacat
     3841 gacgcttcgg cggaacgggc tcgggcagct gggcttccac gtgaagtacg acggcacggt
     3901 ggccgaggtt gaggactatg ggttcgcctg gcaggccggc ctccggcagg gcagccgact
     3961 agtggagatc tgcaaggtgg ccgtggtcac actgacccac gaccagatga tcgacctgct
     4021 gcgcacctct gtcactgtga aggtggtcat catcccgcct tttgaggacg gcactccccg
     4081 gaggggttgg ccggagacct acgacatgaa tacctcggag cccaagacgg agcaggaaag
     4141 catcactcct gggggccggc ccccctaccg cagcaatgct ccctggcagt ggagcgggcc
     4201 cgcatcccat aactctctac cagcctccaa gtgggccact ccaaccactc ccggccatgc
     4261 ccagtccctg agccggcccc tgaagcagac ccccatagtc cccttccggg agtcccagcc
     4321 actgcacagc aagaggcctg tcagcttccc agaaacccct tacacagtat caccagcagg
     4381 ggccgacaga gtccctccct accgacagcc ttctgggagc ttctccaccc ccggttcggc
     4441 cacctacgtg agatacaagc catccccaga aaggtacacg gctgccccac accccctgct
     4501 atctcttgat ccccacttca gccacgatgg gacgtccagc ggcgactcct cttccggcgg
     4561 cctgaccagc caggagagca ccatggaacg ccagaagcca gagcctttgt ggcatgtgcc
     4621 tgcccaggcc aggctctcag ccatagccgg aagcagcggg aacaagcacc cgtccaggca
     4681 ggatgcagca ggcaaagatt cccccaacag gcattccaaa ggagaacctc aatactcaag
     4741 tcattccagc agcaacaccc tctccagcaa cgcatccagc agccacagcg acgaccgctg
     4801 gttcgacccc ctggaccccc tggagccaga gcaagacccc ctctccaagg gtggctctag
     4861 tgacagcggc atcgacacca ccctctacac ctccagccct agctgcatgt ccctggccaa
     4921 ggctccacgg cccgccaagc cacacaagcc ccctggaagt atgggccttt gtggcggggg
     4981 tcgcgaggcc gctgggaggt cccaccacgc agacaggcgg cgggaggtct cccctgcccc
     5041 cgcagttgcc ggccaaagca agggctaccg accgaagctg tactcctccg gctccagcac
     5101 ccccacggga ctggcggggg gcagccgaga cccaccgagg cagcccagtg acatgggctc
     5161 gagggttggc taccccgctc aggtttacaa aactgccagt gcagagactc ctcggccctc
     5221 ccagctggcc cagcccagcc cctttcagct ctccgcctcc gtccccaagt ccttcttctc
     5281 caagcagcct gtacgcaata agcacccaac agggtggaag agaacggagg agcccccacc
     5341 acggccactc cccttcagtg acccaaagaa gcaggtggac acgaacacca aaaatgtctt
     5401 tgggcaaccg aggttgaggg catccctccg agacctccgg tcaccacgga agaactacaa
     5461 atccaccatc gaggatgacc tgaagaaact catcatcatg gacaacctgg ggccagagca
     5521 ggagagagac acgggacagt caccgcagaa gggcctgcag cggacgctgt cggacgagag
     5581 cctgtgcagc gggcgccggg agcccagctt cgccagcccc gctggcctag agccagggct
     5641 gcccagcgac gtgctcttca ccagcacctg cgccttcccg tccagcacgc tgcctgcacg
     5701 ccgccagcac cagcaccccc acccgcccgt cggccccggt gccacccctg ccgccggcag
     5761 cggctttccc gagaagaaat ccaccatctc agcctcggag ctctcgctgg ctgatgggcg
     5821 ggaccgcccc ctgcggcgcc tggaccctgg gctgatgccc ctgcctgaca cagctgctgg
     5881 cctcgagtgg tccagcctgg tgaacgcagc caaggcatac gaagtgcaaa gagccgtctc
     5941 actcttctct ctgaacgacc cggccctgag cccggacatc ccgcctgcac acagtcctgt
     6001 ccacagccac ctgagcctgg agaggggacc cccgaccccc aggaccaccc ctaccatgag
     6061 cgaggagcca cccctggatc tgacaggcaa ggtgtaccag ctggaggtga tgctgaaaca
     6121 gctgcacact gacctgcaga aggagaagca ggacaaggtg gtgctccagt cagaggtggc
     6181 cagcctgcgg cagaacaacc agcggctgca ggaggagtcg caggccgcca gcgagcagct
     6241 gcgcaagttt gcggagatct tctgcaggga gaagaaggag ctcctcgagg actacaaaga
     6301 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     6361 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     6421 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     6481 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     6541 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     6601 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     6661 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6721 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6781 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6841 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6901 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6961 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     7021 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     7081 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     7141 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     7201 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7261 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     7321 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     7381 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     7441 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     7501 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     7561 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     7621 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     7681 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     7741 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     7801 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     7861 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     7921 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     7981 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     8041 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     8101 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     8161 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     8221 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     8281 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     8341 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     8401 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     8461 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     8521 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     8581 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     8641 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     8701 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     8761 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     8821 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     8881 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     8941 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     9001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     9061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     9121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     9181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     9241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     9301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     9361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     9421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     9481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     9541 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     9601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     9661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     9721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     9781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     9841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     9901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     9961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    10021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    10081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    10141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    10201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    10261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    10321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    10381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    10441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    10501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    10561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    10621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    10681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    10741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
