LOCUS       V032191                 7812 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032191
VERSION     V032191
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             874..2532
                     /label="Mcoln3(NM_134160)"
                     /note="Mcoln3(NM_134160)"
                     /gene="Mcoln3"
     CDS             2569..3279
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     polyA_signal    3379..3603
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3649..4077
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4091..4420
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4487..5281
                     /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    5455..5576
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5625..5641)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5649..5665
                     /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(5673..5703)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5718..5739
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6027..6612)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6783..7643)
                     /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(7644..7748)
                     /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 ggctagcgcc accatggcaa atcccgaggt gctggttagc
      901 agctgcagag ctcgccaaga tgaaagcccc tgcactttcc acccgagctc gtccccgtca
      961 gagcagcttc tcttagaaga ccagatgagg cggaaactca agttcttttt tatgaatcct
     1021 tgtgagaagt tctgggctcg gggtaggaag ccatggaaac ttgccataca gattctgaaa
     1081 atcgcgatgg tgactatcca gctggttctg tttggactaa gtaaccagat ggtagtagct
     1141 ttcaaagagg agaacactat agccttcaaa cacctcttcc taaagggcta catggatcga
     1201 atggacgaca cctatgcagt gtacactcag agtgaagtgt atgaccagat catctttgca
     1261 gtgacccagt acttgcagct tcagaacatc tccgtgggca atcacgctta tgagaacaag
     1321 gggactaagc agtcggcgat ggcaatctgt cagcacttct acaggcaagg aaccatctgc
     1381 cccgggaacg acacctttga catcgatcca gaagttgaaa cagaatgttt ccttgtagag
     1441 ccagatgaag cttcccacct tggaacgcct ggagaaaata aactcaacct gagcctggac
     1501 ttccacagac ttctgacggt ggagctccag tttaagctca aagccatcaa tctgcagaca
     1561 gttcgacacc aggagcttcc tgactgttac gactttacgc tgactataac attcgacaac
     1621 aaggctcaca gtggaagaat caaaataagc ttagacaacg acatttctat caaagaatgc
     1681 aaagactggc atgtgtctgg atcaattcag aagaacacac actacatgat gatctttgat
     1741 gcctttgtca ttctgacctg cttggcctca ctggtgctgt gtgccaggtc tgtgattagg
     1801 ggtcttcagc ttcagcagga gtttgtcaac ttcttccttc ttcactacaa gaaggaagtt
     1861 tcggcctctg atcagatgga gttcatcaac gggtggtaca ttatgatcat cattagtgac
     1921 atattgacaa tcgttggatc agttctgaaa atggaaatcc aagccaagag tctcacaagc
     1981 tatgatgtct gcagcatact tctcgggacg tcaactatgc tcgtgtggct tggagttatc
     2041 cgatacctgg gtttctttgc gaagtacaat ctccttattc tgaccctcca ggcagcgctg
     2101 cccaacgtca tgaggttctg ttgctgcgct gctatgatct atctaggcta ttgcttttgc
     2161 ggatggattg tgctgggccc ttaccatgag aagttccgtt ccctgaacag ggtctccgag
     2221 tgcctgttct cgctgataaa cggagacgat atgttttcca catttgcgaa aatgcagcag
     2281 aagagttacc tggtgtggct gttcagccga gtctacctgt actcgttcat cagcctcttc
     2341 atttacatga ttctgagcct tttcatcgcg ctcatcacag acacatacga aacaattaag
     2401 cactaccagc aagatggctt cccagagacg gaacttcgaa agtttatagc ggaatgcaaa
     2461 gacctcccca actccggaaa atacagatta gaagatgacc ctccgggttc tttactctgc
     2521 tgctgcaaaa agaagcttgg cggtggaggc agtggtggag gcggtagcat ggtgagcaag
     2581 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     2641 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2701 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     2761 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     2821 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2881 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     2941 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     3001 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     3061 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     3121 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     3181 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     3241 cgccactcca ccggcggcat ggacgagctg tacaagtagg gatccactag tccagtgtgg
     3301 tggaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa
     3361 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3421 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3481 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3541 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3601 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3661 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3721 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3781 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3841 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3901 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3961 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4021 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4081 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4141 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4201 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4261 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4321 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4381 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4441 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4501 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4561 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4621 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4681 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4741 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4801 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4861 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4921 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4981 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5041 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5101 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5161 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5221 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5281 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5341 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5401 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5461 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5521 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5581 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5641 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5701 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5761 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5821 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5881 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5941 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6001 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6061 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6121 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6181 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6241 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6301 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6361 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6421 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6481 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6541 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6781 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6841 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6901 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6961 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7021 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7081 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7141 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7201 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7261 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7321 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7381 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7441 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7501 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7561 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7621 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7741 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7801 atcccctatg gt
//
