LOCUS       V037701                 9063 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037701
VERSION     V037701
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             700..3789
                     /label="PPP1R12A(NM_002480)"
                     /note="PPP1R12A(NM_002480)"
                     /gene="PPP1R12A"
     CDS             3796..3819
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    3839..4427
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4493..4541
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4743..5171
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5185..5514
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5581..6375
                     /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    6551..6672
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6721..6737)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6745..6761
                     /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(6769..6799)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6814..6835
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7123..7711)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7882..8742)
                     /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(8743..8847)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattca tgaagatggc ggacgcgaag
      721 cagaagcgga acgagcagct gaaacgctgg atcggctccg agacggacct cgagcctccg
      781 gtggtgaagc gccagaagac caaggtgaag ttcgacgatg gcgccgtctt cctggctgct
      841 tgctccagcg gcgacacgga cgaggtcctc aagctgctgc accgcggcgc cgacatcaat
      901 tacgccaatg tggacggact cactgccctg caccaggctt gcattgatga caatgttgat
      961 atggtgaagt ttctggtaga aaatggagca aatattaatc aacctgataa tgaaggctgg
     1021 ataccactac atgcagcagc ttcctgtgga tatcttgata ttgcagagtt tttgattggt
     1081 caaggagcac atgtaggggc tgtcaacagt gaaggagata cacctttaga tattgcggag
     1141 gaggaggcaa tggaagagct acttcaaaat gaagttaatc ggcaaggggt tgatatagaa
     1201 gcagctcgaa aggaagaaga acggatcatg cttagagatg ccaggcagtg gctaaatagt
     1261 ggtcatataa atgatgtccg gcatgcaaaa tctggaggta cagcacttca cgttgcagct
     1321 gctaaaggct atacggaagt tttaaaactt ttaatacagg caggctatga tgttaatatt
     1381 aaagactatg atggctggac acctcttcat gctgcagctc attggggtaa agaagaagca
     1441 tgtcgaattt tagtggacaa tctgtgtgat atggagatgg tcaacaaagt gggccaaaca
     1501 gcctttgatg tagcagatga agacatttta ggatatttag aagagttgca aaagaaacaa
     1561 aatctgctcc atagtgaaaa acgggacaag aaatctccac taattgaatc aacagcaaat
     1621 atggacaata atcagtcaca gaagaccttt aaaaacaaag agacgttgat tattgaacca
     1681 gagaaaaatg catcccgtat tgaatctctg gaacaagaaa aggttgatga agaagaagaa
     1741 ggaaagaagg atgagtctag ctgctctagt gaagaagatg aggaagatga ctcggaatca
     1801 gaagctgaaa cagataagac aaaacccctg gcttctgtaa ctaatgccaa cacttctagt
     1861 acacaagcag ctcctgtagc tgttacaaca cctactgtgt catcaggtca agcaacacct
     1921 acatcaccta ttaaaaagtt tccaaccaca gctacaaaaa tttctcccaa agaagaagag
     1981 agaaaagatg agtctcctgc aacttggagg ttaggactta gaaagacggg cagctatggt
     2041 gcacttgctg aaatcacagc atctaaagag ggtcagaaag aaaaagatac tgcaggtgtt
     2101 acacgttcag cttcaagtcc cagactttcc tcctctttgg ataataaaga aaaggagaaa
     2161 gatagtaaag gaactaggct tgcatatgtt gcacctacaa taccaagacg actagccagt
     2221 acatctgaca ttgaagagaa agaaaacaga gattcttcaa gtttgcgaac aagtagttca
     2281 tatacaagga gaaaatggga agatgatctt aaaaaaaata gctcagttaa tgaaggatca
     2341 acgtatcata aaagttgctc ctttggtaga agacaagatg atttgattag ttctagtgtt
     2401 ccaagcacca catcaacacc aacagttacc tctgcagctg ggcttcagaa aagcctgctt
     2461 tccagcacaa gcactactac aaagattaca acgggttctt cctcagcagg cacacaaagc
     2521 agtacctcaa atcgtttgtg ggctgaggat agtactgaga aagaaaagga cagtgttcct
     2581 acggcagtga ccattcctgt tgctccaact gttgtaaatg ctgcagcttc taccacaacc
     2641 ctgactacaa ctactgctgg cactgtctcc tccacaacag aggtcaggga gagacgcaga
     2701 tcatacctca ctcctgttag ggatgaagag tctgaatccc aaagaaaagc aagatctaga
     2761 caagcaagac aatctagaag atcaacacag ggagtgacat taactgatct tcaagaagct
     2821 gagaaaacaa taggaagaag tcgttctacc cgaaccagag aacaagaaaa tgaagaaaaa
     2881 gaaaaagagg aaaaagagaa acaagataaa gagaaacaag aagaaaagaa ggagtcagaa
     2941 acatctagag aagatgaata taaacaaaag tactccagaa cgtatgatga gacttaccag
     3001 cgttataggc cagtatcaac ttcaagttca accactccat cctcttcact ttctactatg
     3061 agcagttcac tgtatgcttc aagtcaacta aacaggccaa atagtcttgt aggcataact
     3121 tctgcttact ccagaggaat aacaaaagaa aatgaaagag agggagaaaa aagagaagag
     3181 gagaaagaag gagaagataa atcacaacct aaatcaatca gagaacgacg acgaccaaga
     3241 gagaaaagaa gatctacagg agtttcattt tggacacaag atagtgatga aaatgaacaa
     3301 gaacaacaat cagacacaga agagggatcc aataagaaag aaactcagac ggattccatt
     3361 tctagatatg aaaccagttc tacatcagct ggtgatcgat atgattcctt gctgggtcgc
     3421 tctggatcat acagttactt agaagaaaga aaaccttaca gcagcaggct agaaaaggat
     3481 gactcaactg actttaaaaa gctttatgaa caaattctag ctgaaaatga aaagctgaag
     3541 gcacagctac atgatacaaa tatggaacta acagatctta aattacagtt ggaaaaggcc
     3601 acccagagac aagaaagatt tgctgataga tcactgttgg aaatggaaaa aagggaacga
     3661 agagctctag aaagaagaat atctgaaatg gaagaagagc tcaaaatgtt accagaccta
     3721 aaagcagaca accagaggct aaaggatgaa aatggggcct tgatcagagt tataagcaaa
     3781 ctttccaaac tcgagcacca tcaccatcac catcaccatt agggcccgat atctcgacaa
     3841 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3901 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3961 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4021 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4081 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4141 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4201 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4261 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4321 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4381 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     4441 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4501 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4561 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4621 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4681 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     4741 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4801 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4861 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4921 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4981 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5041 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5101 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5161 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5221 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5281 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5341 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5401 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5461 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5521 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5581 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5641 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5701 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5761 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5821 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5881 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5941 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6001 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6061 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6121 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6181 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6241 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6301 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6361 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6421 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6481 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6541 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6661 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6721 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6781 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6841 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6901 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     7321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7561 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7741 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7801 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7861 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7921 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7981 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8041 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8101 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8161 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8221 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8281 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8341 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8401 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8461 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8521 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8581 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8641 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8701 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8761 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8821 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8881 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     8941 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     9001 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     9061 agg
//
