LOCUS       V021946                 6888 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021946
VERSION     V021946
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             898..924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             940..2376
                     /label="PPM1B(NM_002706)"
                     /note="PPM1B(NM_002706)"
                     /gene="PPM1B"
     promoter        complement(2440..2458)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2484..2708
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2754..3182
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3196..3526
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3593..4387
                     /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    4561..4682
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4731..4747)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4755..4771
                     /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(4779..4809)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4824..4845
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5133..5721)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5892..6752)
                     /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(6753..6857)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ggagacccgc caccatgtac
      901 ccatacgacg tcccagacta cgctggaagc ttggtaccga tgggtgcatt tttggataaa
      961 cccaaaactg aaaaacataa tgctcatggt gctgggaatg gtttacgtta tggcctgagc
     1021 agcatgcaag gatggagagt ggaaatggaa gatgcacaca cagctgttgt aggtattcct
     1081 cacggcttgg aagactggtc attttttgca gtttatgatg gtcatgctgg atcccgagtg
     1141 gcaaattact gctcaacaca tttattagaa cacatcacta ctaacgaaga ctttagggca
     1201 gctggaaaat caggatctgc tcttgagctt tcagtggaaa atgttaagaa tggtatcaga
     1261 actggatttt tgaaaattga tgaatacatg cgtaactttt cagacctcag aaacgggatg
     1321 gacaggagtg gttcaactgc agtgggagtt atgatttcac ctaagcatat ctactttatc
     1381 aactgtggtg attcacgtgc tgttctgtat aggaatggac aagtctgctt ttctacccag
     1441 gatcacaaac cttgcaatcc aagggaaaag gagcgaatcc aaaatgcagg aggcagcgtg
     1501 atgatacaac gtgttaatgg ttcattagca gtatctcgtg ctctggggga ctatgattac
     1561 aagtgtgttg atggcaaggg cccaacagaa caacttgttt ctccagagcc tgaggtttat
     1621 gaaattttaa gagcagaaga ggatgaattt atcatcttgg cttgtgatgg gatctgggat
     1681 gttatgagta atgaggagct ctgtgaatat gttaaatcta ggcttgaggt atctgatgac
     1741 ctggaaaatg tgtgcaattg ggtagtggac acttgtttac acaagggaag tcgagataac
     1801 atgagtattg tactagtttg cttttcaaat gctcccaagg tctcagatga agcggtgaaa
     1861 aaagattcag agttggataa gcacttggaa tcacgggttg aagagattat ggagaagtct
     1921 ggcgaggaag gaatgcctga tcttgcccat gtcatgcgca tcttgtctgc agaaaatatc
     1981 ccaaatttgc ctcctggggg aggtcttgct ggcaagcgta atgttattga agctgtttat
     2041 agtagactga atccacatag agaaagtgat ggggcctccg atgaagcaga ggaaagtgga
     2101 tcacagggaa aattggtgga agctctcagg caaatgagaa ttaatcatag gggaaactac
     2161 cgacaacttc tggaggagat gctgactagt tacaggctag ctaaagtaga gggagaagaa
     2221 agccctgctg aaccagctgc cacagctact tcttcgaaca gtgatgctgg aaacccagtg
     2281 acaatgcagg aaagccatac tgaatcagaa agtggtcttg ctgaattaga cagctctaat
     2341 gaagatgcag ggacaaagat gagtggtgaa aaaatatgag aattctgcag atatccatca
     2401 cactggcggc cgctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     2461 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2521 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2581 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2641 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2701 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2761 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2821 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2881 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     2941 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3001 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3061 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3121 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3181 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg
     3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3481 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3541 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3601 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3661 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3721 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3781 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3841 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3901 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3961 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4021 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4081 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4141 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4201 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4261 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4321 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4381 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4441 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4501 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4561 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4621 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4681 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4741 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4801 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4861 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4921 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4981 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5041 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5101 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5161 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5221 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5281 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     5341 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5401 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5461 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5521 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5581 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5641 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5701 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5761 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5821 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5881 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5941 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6001 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6061 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6121 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6181 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6241 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6301 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6361 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6421 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6481 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6541 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6601 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6661 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6721 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6781 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6841 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
