LOCUS       V037464                 8102 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037464
VERSION     V037464
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             1883..3622
                     /label="MCOLN1(NM_020533)"
                     /note="MCOLN1(NM_020533)"
                     /gene="MCOLN1"
     polyA_signal    3669..3893
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3939..4367
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4381..4710
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4777..5571
                     /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    5745..5866
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5915..5931)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5939..5955
                     /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(5963..5993)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6008..6029
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6317..6902)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7073..7933)
                     /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(7934..8038)
                     /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 ttcaaacacc ctatggaacc attagcttgt aattgggagg actgtgataa
      961 gcttggagat gatacatgtt ccatagtgaa ccatattaat tgtatccttc tagagaacgt
     1021 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     1081 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1141 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccatac tttcgaaaga
     1201 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1261 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga
     1321 ggagctgttc accggggtgg tgcccatcca ggtcgagctg gacggcgacg taaacggcca
     1381 caagttcagc gtgcgcggcg agggtgaggg cgatgccacc aacggcaagc tgaccctgaa
     1441 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1501 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1561 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcac
     1621 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1681 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct
     1741 gccgaaaaac aatgacttaa aatgcaaatg gaaagaatgt cctgagtctt gtagctcact
     1801 aatggaccta caaagacatc ttttgaagga tcatgtctct caagatggtg gaggcggatc
     1861 aggaggcggt ggatctgaat tcatgacagc cccggcgggt ccgcgcggct cagagaccga
     1921 gcggcttctg acccccaacc ccgggtatgg gacccaggcg gggccttcac cggcccctcc
     1981 gacaccccca gaagaggaag accttcgccg tcgtctcaaa tactttttca tgagtccctg
     2041 cgacaagttt cgagccaagg gccgcaagcc ctgcaagctg atgctgcaag tggtcaagat
     2101 cctggtggtc acggtgcagc tcatcctgtt tgggctcagt aatcagctgg ctgtgacatt
     2161 ccgggaagag aacaccatcg ccttccgaca cctcttcctg ctgggctact cggacggagc
     2221 ggatgacacc ttcgcagcct acacgcggga gcagctgtac caggccatct tccatgctgt
     2281 ggaccagtac ctggcgttgc ctgacgtgtc actgggccgg tatgcgtatg tccgtggtgg
     2341 gggtgaccct tggaccaatg gctcagggct tgctctctgc cagcggtact accaccgagg
     2401 ccacgtggac ccggccaacg acacatttga cattgatccg atggtggtta ctgactgcat
     2461 ccaggtggat ccccccgagc ggccccctcc gccccccagc gacgatctca ccctcttgga
     2521 aagcagctcc agttacaaga acctcacgct caaattccac aagctggtca atgtcaccat
     2581 ccacttccgg ctgaagacca ttaacctcca gagcctcatc aataatgaga tcccggactg
     2641 ctataccttc agcgtcctga tcacgtttga caacaaagca cacagtgggc ggatccccat
     2701 cagcctggag acccaggccc acatccagga gtgtaagcac cccagtgtct tccagcacgg
     2761 agacaacagc ttccggctcc tgtttgacgt ggtggtcatc ctcacctgct ccctgtcctt
     2821 cctcctctgc gcccgctcac tccttcgagg cttcctgctg cagaacgagt ttgtggggtt
     2881 catgtggcgg cagcggggac gggtcatcag cctgtgggag cggctggaat ttgtcaatgg
     2941 ctggtacatc ctgctcgtca ccagcgatgt gctcaccatc tcgggcacca tcatgaagat
     3001 cggcatcgag gccaagaact tggcgagcta cgacgtctgc agcatcctcc tgggcacctc
     3061 gacgctgctg gtgtgggtgg gcgtgatccg ctacctgacc ttcttccaca actacaatat
     3121 cctcatcgcc acactgcggg tggccctgcc cagcgtcatg cgcttctgct gctgcgtggc
     3181 tgtcatctac ctgggctact gcttctgtgg ctggatcgtg ctggggccct atcatgtgaa
     3241 gttccgctca ctctccatgg tgtctgagtg cctgttctcg ctcatcaatg gggacgacat
     3301 gtttgtgacg ttcgccgcca tgcaggcgca gcagggccgc agcagcctgg tgtggctctt
     3361 ctcccagctc tacctttact ccttcatcag cctcttcatc tacatggtgc tcagcctctt
     3421 catcgcgctc atcaccggcg cctacgacac catcaagcat cccggcggcg caggcgcaga
     3481 ggagagcgag ctgcaggcct acatcgcaca gtgccaggac agccccacct ccggcaagtt
     3541 ccgccgcggg agcggctcgg cctgcagcct tctctgctgc tgcggaaggg acccctcgga
     3601 ggagcattcg ctgctggtga attgactcga gtctagaggg cccgtttaaa cccgctgatc
     3661 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3721 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3781 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3841 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3901 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3961 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4021 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4081 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4141 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4201 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4261 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4321 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4381 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4441 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4501 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4561 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4621 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4681 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4741 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4801 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4861 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4921 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4981 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5041 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5101 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5161 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5221 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5281 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5341 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5401 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5461 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5521 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5581 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5641 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5701 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5761 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5821 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5881 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5941 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6001 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6061 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6121 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6181 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6241 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6301 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6361 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6421 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6481 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6541 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6601 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6661 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6721 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6781 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6841 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6901 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6961 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7021 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7081 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7141 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7201 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7261 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7321 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7381 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7441 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7501 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7561 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7621 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7681 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7741 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7801 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7861 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7921 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7981 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8041 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8101 gt
//
