LOCUS       V023474                 8042 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023474
VERSION     V023474
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..3532
                     /label="Mlxipl(NM_021455)"
                     /note="Mlxipl(NM_021455)"
                     /gene="Mlxipl"
     CDS             3539..3604
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3642..3866
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3912..4340
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4354..4683
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4750..5544
                     /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    5718..5839
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5888..5904)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5912..5928
                     /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(5936..5966)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5981..6002
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6290..6875)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7046..7906)
                     /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(7907..8011)
                     /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 atggcgcgcg cgctggcgga
      961 tctatccgtg aacttgcagg tcccccgggt cgtccctagc ccggactcgg actcggatac
     1021 ggacttggag gatccgagtc cccggcgcag cgcgggtggc ctgcatcgat cacaggtcat
     1081 ccacagcgga cacttcatgg tgtcttcgcc gcacagcgac tcgctgaccc ggcgacgcga
     1141 ccaggagggg cccgtggggc tcgccgactt cgggccgcgc agcatcgatc cgacactcac
     1201 ccacctcttc gagtgcttga gcctggctta cagtggcaag ctggtctctc ccaagtggaa
     1261 gaacttcaaa ggcctcaagt tgctatgccg ggacaagatc cggctgaaca acgccatctg
     1321 gagagcctgg tacattcagt atgtgcaacg gaggaagagc ccagtgtgtg gtttcgtgac
     1381 ccctctgcag gggtctgaag cagatgagca ccggaaacct gaggctgtca tcctggaggg
     1441 taattactgg aagcggcgca tcgaggtggt gatgcgcgaa taccacaagt ggagaatcta
     1501 ctacaaaaag cggctccgta agtccagcag ggaaggggat ttcctggctc ccaagcaggt
     1561 ggaaggtggg tggccgccac cagagcgatg gtgcgaacag ctcttctcca gcgtggtgcc
     1621 tgtgctgctt gggggctccg aggaggagcc tgggggccgg cagcttctag acctggactg
     1681 cttcttgtcc gatatctccg acacactctt caccatgaca cagcccagtc cttcgtccct
     1741 gcagctgccc ccagaagatg cttatgttgg caatgctgac atgatccagc cagacctgac
     1801 gccgctgcag cccagcctag atgacttcat ggagatatca gatttcttca ccaattaccg
     1861 cccccctcag acacccacat cttcaaacta catagagtcc cccagctttg gccccatggc
     1921 tgactccctc ttcagcagtg ggatcctggc cccagagatg ccatctccag cctcgtcttc
     1981 ctcctcctca gggatgaccc ctcactcagg gaatacacgc ctacaggctc ggaacagctg
     2041 ctctggaccc ttggatccca acccctttct gagctctgaa ttccttcttc ctgaagaccc
     2101 taagaccaag atcccacctg ctcctggacc gacccctctc cttccattcc ccacccctgt
     2161 caaggtgcat ggcttagagc cctgcacccc ctctcccttc cccacaatgg ctccgccccc
     2221 ctctttgttg ccagaagaat ctctcctctc tgccaggttc ccattcacct cagcccctcc
     2281 tgctccagga gtgtctacgc ttcctgcacc cacaaccttt gtccccaccc cacagcccgg
     2341 ccctggtccc gtccccttct ctgtagacca tctgccccat gggtacctgg aacccgtctt
     2401 tgggcctcac ttcactgtgc ctcagggtat gcaacccagg tgcaagccct cctccccatc
     2461 ccctggggga caaaaagcaa gtccacccac cttggcctct gccaccgcca gtcccactgc
     2521 cactgccact gccagagaca acaacccctg tcttacacag ctgctcaggg cagccaagcc
     2581 tgagcaagca ctggagcctc cgactatgcc cggtaccctc ctccggcctc cagagtcccc
     2641 gcaggataca gtttctgaaa tcccccgtgc ccgtgccttt ttccccccaa tcccggcccc
     2701 tacaccaccc cggccacctc caggcccagc cacattggct cctcccaggt ccctggttgt
     2761 ccccaaagca gagcggctct cgcctccagc ctccagcggc agtgagcggc gactatcagg
     2821 ggatctcaac tccatacaac cctcgggggc actgagtgtc cacctgtctc cccctcaaac
     2881 tgtcctaagc cggggtcgtg tagacaacaa caagatggag aaccgacgta tcacacacat
     2941 ctccgcggag cagaagaggc ggttcaatat taagctagga ttcgacaccc tgcacggact
     3001 tgtcagcacg ctcagtgccc agcccagcct caaggtgagc aaagcaacca cgcttcagaa
     3061 gacagcggag tacatcctga tgctgcagca ggaacgggca gctatgcagg aggaggcgca
     3121 gcagctgcgg gatgaaatag aggagctcaa cgctgccatc aacttgtgcc agcagcagct
     3181 accggccacc ggggtgccca tcacacatca gcgctttgac cagatgcggg acatgtttga
     3241 tgactatgtc cggacgcgca cgctgcacaa ctggaagttc tgggtgttca gcatcctcat
     3301 ccgaccttta tttgaatcct tcaatgggat ggtgtctacc gccagcttgc acagcctccg
     3361 ccagacctca ctggcctggc tggaacagta ctgttcccta cctgctctcc ggccaactgt
     3421 cctgaattcc cttcgccaac tcagcacttc cacaagcatc ctgactgacc ccagccttgt
     3481 gcctgagcaa gccacacggg cagtcacaga gggcaccctg gggagaccat tactcgagga
     3541 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3601 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3661 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3721 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3781 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3841 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3901 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4621 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4681 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4741 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4801 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4861 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4921 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4981 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5041 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5101 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5161 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5221 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5281 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5341 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5401 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5461 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5521 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5581 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5641 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5701 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5761 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5821 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5881 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5941 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6001 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6061 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6121 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6181 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6241 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6301 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6361 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6421 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6481 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6541 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6601 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6661 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6721 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6781 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8041 tc
//
