LOCUS       V035648                 8093 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035648
VERSION     V035648
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             884..1591
                     /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"
     CDS             1628..3613
                     /label="Mmp2(NM_008610)"
                     /note="Mmp2(NM_008610)"
                     /gene="Mmp2"
     polyA_signal    3660..3884
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3930..4358
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4372..4701
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4768..5562
                     /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    5736..5857
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5906..5922)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5930..5946
                     /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(5954..5984)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5999..6020
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6308..6893)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7064..7924)
                     /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(7925..8029)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg gaggcacgag tggcctgggg agcgctggcc ggacctctgc gggttctctg
     1681 cgtcctgtgc tgcctgttgg gccgcgccat cgctgcacca tcgcccatca tcaagttccc
     1741 cggcgatgtc gcccctaaaa cagacaaaga gttggcagtg caatacctga acactttcta
     1801 tggctgcccc aaggagagtt gcaacctctt tgtgctgaaa gataccctca agaagatgca
     1861 gaagttcttt gggctgcccc agacaggtga ccttgaccag aacaccatcg agaccatgcg
     1921 gaagccaaga tgtggcaacc cagatgtggc caactacaac ttcttccccc gcaagcccaa
     1981 gtgggacaag aaccagatca catacaggat cattggttac acacctgacc tggaccctga
     2041 aaccgtggat gatgcttttg ctcgggcctt aaaagtatgg agcgacgtca ctccgctgcg
     2101 cttttctcga atccatgatg gggaggctga catcatgatc aactttggac gctgggagca
     2161 tggagatgga tacccatttg atggcaagga tggactcctg gcacatgcct ttgccccggg
     2221 cactggtgtt gggggagatt ctcactttga tgatgatgag ctgtggaccc tgggagaagg
     2281 acaagtggtc cgcgtaaagt atgggaacgc tgatggcgag tactgcaagt tccccttcct
     2341 gttcaacggt cgggaataca gcagctgtac agacactggt cgcagtgatg gcttcctctg
     2401 gtgctccacc acatacaact ttgagaagga tggcaagtat ggcttctgcc cccatgaagc
     2461 cttgtttacc atgggtggca atgcagatgg acagccctgc aagttcccgt tccgcttcca
     2521 gggcacctcc tacaacagct gtaccaccga gggccgcacc gatggctacc gctggtgtgg
     2581 caccaccgag gactatgacc gggataagaa gtatggattc tgtcccgaga ccgctatgtc
     2641 cactgtgggt ggaaattcag aaggtgcccc atgtgtcttc cccttcactt tcctgggcaa
     2701 caagtatgag agctgcacca gcgccggccg caacgatggc aaggtgtggt gtgcgaccac
     2761 aaccaactac gatgatgacc ggaagtgggg cttctgtcct gaccaaggat atagcctatt
     2821 cctcgtggca gcccatgagt tcggccatgc catggggctg gaacactctc aggaccctgg
     2881 agctctgatg gccccgatct acacctacac caagaacttc cgattatccc atgatgacat
     2941 caaggggatc caggagctct atgggccctc ccccgatgct gatactgaca ctggtactgg
     3001 ccccacacca acactgggac ctgtcactcc ggagatctgc aaacaggaca ttgtctttga
     3061 tggcatcgct cagatccgtg gtgagatctt cttcttcaag gaccggttta tttggcggac
     3121 agtgacacca cgtgacaagc ccacaggtcc cttgctggtg gccacattct ggcctgagct
     3181 cccagaaaag attgacgctg tgtatgaggc cccacaggag gagaaggctg tgttcttcgc
     3241 agggaatgag tactgggtct attctgctag tactctggag cgaggatacc ccaagccact
     3301 gaccagcctg gggttgcccc ctgatgtcca gcaagtagat gctgccttta actggagtaa
     3361 gaacaagaag acatacatct ttgcaggaga caagttctgg agatacaatg aagtgaagaa
     3421 gaaaatggac cccggtttcc ctaagctcat cgcagactcc tggaatgcca tccctgataa
     3481 cctggatgcc gtcgtggacc tgcagggtgg tggtcatagc tacttcttca agggtgctta
     3541 ttacctgaag ctggagaacc aaagtctcaa gagcgtgaag tttggaagca tcaaatcaga
     3601 ctggctgggc tgctgaggat cctctagagg gcccgtttaa acccgctgat cagcctcgac
     3661 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3721 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3781 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3841 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3901 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3961 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4021 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4081 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4141 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4201 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4261 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4321 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4381 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4441 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4501 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4561 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4621 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4681 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4741 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4801 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4861 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4921 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4981 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5041 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5101 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5161 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5221 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5281 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5341 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5401 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5461 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5521 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5581 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5641 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5701 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5761 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5821 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5881 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5941 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6001 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6061 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6121 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6721 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6841 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6901 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6961 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7021 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7081 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7141 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7201 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7261 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7321 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7381 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7441 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7501 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7561 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7621 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7681 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7741 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7801 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7861 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7921 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7981 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8041 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
