LOCUS       V044514                 8098 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044514
VERSION     V044514
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..3588
                     /label="MIOS(NM_001370076)"
                     /note="MIOS(NM_001370076)"
                     /gene="MIOS"
     CDS             3595..3660
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3698..3922
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3968..4396
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4410..4739
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4806..5600
                     /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    5774..5895
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5944..5960)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5968..5984
                     /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(5992..6022)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6037..6058
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6346..6931)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7102..7962)
                     /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(7963..8067)
                     /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 accatgagcg gtaccaaacc tgatatttta tgggcaccac accatgttga tagatttgtt
     1021 gtgtgtgact cagaactaag tctttatcat gtggaatcta ctgtgaattc agaactcaaa
     1081 gctggatctt tacgtttatc tgaagactct gcagctacat tactgtcaat aaattcagat
     1141 acaccctata tgaaatgtgt tgcctggtat cttaattatg atcctgaatg tctgctggca
     1201 gttggacaag caaatggtcg agttgtactt acaagccttg gtcaagatca taactcaaag
     1261 ttcaaagatt tgataggaaa agagtttgtt ccaaaacatg cacgacaatg taataccctt
     1321 gcctggaatc cactggatag taactggcta gctgctggtt tagataagca cagagctgac
     1381 ttttcagtgc taatatggga tatctgcagc aaatatactc ctgatatagt tcccatggaa
     1441 aaagtgaaac tttcagcagg tgaaactgaa acaacattat tagtaacaaa accactttat
     1501 gagttaggac agaatgatgc ttgtctgtct ctttgttggc ttccacgaga ccagaaactt
     1561 ctccttgctg gtatgcatcg taacctagct atatttgatc ttcggaatac aagccaaaag
     1621 atgttcgtaa atacaaaagc tgttcagggt gtgacggtag acccatattt ccacgatcgt
     1681 gttgcttcct tctatgaagg tcaggttgca atatgggatc ttagaaaatt tgagaagcca
     1741 gttttgacat tgactgagca accaaaaccc ttaacaaaag tagcatggtg tcccactagg
     1801 actggtctac ttgccacttt aacaagggat agtaatatta ttagattgta tgatatgcag
     1861 catacaccca ctcccattgg ggatgaaact gaacccacaa taattgaaag aagtgtgcaa
     1921 ccttgtgaca attacattgc ttcctttgcg tggcatccaa caagtcaaaa tcgaatgata
     1981 gttgtaactc ccaaccgaac aatgtcagac ttcactgttt ttgaaaggat atctcttgcc
     2041 tggagcccaa ttacatcttt aatgtgggct tgtggtcgtc atttatatga atgtacggaa
     2101 gaagaaaatg ataattcttt agaaaaagat atagcaacga agatgcgtct tcgggcttta
     2161 tcaaggtatg gacttgatac agagcaggtg tggaggaacc acattttagc tggaaatgaa
     2221 gatccacagc tcaagtcact ctggtatact ctgcacttta tgaagcaata cacagaagat
     2281 atggatcaga aatctccagg caacaaagga tcattggttt atgcaggaat taaatcaatt
     2341 gtaaagtcat cgttgggaat ggtggaaagc agcagacata attggagtgg gttggataag
     2401 caaagtgata ttcaaaattt aaatgaagag agaatcttag ctttacagct ttgtgggtgg
     2461 ataaagaaag gaacggatgt agacgtgggg ccatttttga actcccttgt acaagaaggg
     2521 gaatgggaaa gagctgctgc tgtggcattg ttcaacttgg atattcgccg agcaatccaa
     2581 atcctgaatg aaggggcatc ttctgaaaaa ggagatctga atctcaatgt ggtagcaatg
     2641 gctttatcgg gttatacgga tgagaagaac tccctttgga gagaaatgtg tagcacactg
     2701 cgattacagc taaataaccc gtatttgtgt gtcatgtttg catttctgac aagtgaaaca
     2761 ggatcttacg atggagtttt gtatgaaaac aaagttgcag tacgtgacag agtggcattt
     2821 gcttgtaaat tccttagtga tactcagtta aatagataca tcgaaaagtt gaccaatgaa
     2881 atgaaagagg ctggaaattt ggaaggaatt ttgcttacag gccttactaa agatggagtg
     2941 gacttaatgg agagttatgt tgatagaact ggagatgttc aaacagcaag ttactgtatg
     3001 ttacagggtt cacctttaga tgttcttaaa gatgaaaggg ttcagtactg gattgagaat
     3061 tatagaaatt tattagatgc ctggaggttt tggcataaac gagctgaatt tgatattcac
     3121 aggagtaagt tggatcccag ttccaagcct ttagcacaag tttttgtgag ttgcaatttc
     3181 tgtggcaagt caatctccta cagctgttca gctgtgcctc atcagggcag aggttttagt
     3241 cagtatggtg tgagtggctc accaacgaaa tctaaagtca caagttgtcc tggctgtcga
     3301 aaaccacttc ctcgatgtgc gctttgtctc attaatatgg gaacaccagt ttctagctgt
     3361 cctggaggaa ccaaatcaga tgaaaaagtg gacttgagca aggacaaaaa attagcccaa
     3421 tttaacaact ggtttacatg gtgtcataat tgcaggcacg gtggacatgc tggacatatg
     3481 cttagttggt tcagggacca tgcagagtgc cctgtgtcgg catgcacgtg taaatgtatg
     3541 cagttggata caacagggaa tctggtacct gcagagactg tccagccact cgaggactac
     3601 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     3661 tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3721 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3781 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3841 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3901 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3961 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4021 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4081 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4141 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4201 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4261 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4321 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4381 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4441 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4501 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4561 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4621 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4681 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4741 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4801 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4861 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4921 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4981 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5041 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5101 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5161 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5221 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5281 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5341 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5401 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5461 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5521 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5581 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5641 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5701 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5761 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5881 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5941 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6001 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6061 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6181 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6241 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6301 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6361 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6421 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6481 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6541 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6601 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6661 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6721 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6781 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6841 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     6901 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6961 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7021 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7081 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7141 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7201 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7261 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7321 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7381 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7441 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7501 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7561 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7621 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7681 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7741 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7801 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7861 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7921 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7981 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8041 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
