LOCUS       V025415                 7001 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025415
VERSION     V025415
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..2491
                     /label="Acvrl1(NM_001277255)"
                     /note="Acvrl1(NM_001277255)"
                     /gene="Acvrl1"
     CDS             2498..2563
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2601..2825
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2871..3299
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3313..3642
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3709..4503
                     /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    4677..4798
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4847..4863)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4871..4887
                     /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(4895..4925)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4940..4961
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5249..5834)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6005..6865)
                     /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(6866..6970)
                     /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 atggcaggaa atctcaccac
      961 atctcttctc ctatctccaa ggaccatgac cttggggagc ttcagaaggg gccttttgat
     1021 gctgtcggtg gcctttggcc taaccagggg agacttggcg aagccttcca agctggtgaa
     1081 ctgcacttgt gagagcccac actgcaagag accattctgc caggggtcat ggtgcacagt
     1141 ggtgctggtt cgggagcagg gcaggcaccc ccaggtctat cggggctgcg ggagcctgaa
     1201 ccaggagctc tgcttgggac gtcccacaga gtttctgaac catcactgct gctatagatc
     1261 cttctgcaac cacaacgtgt ctctgatgct ggaggccacc caaactcctt cggaggagcc
     1321 agaagttgat gcccatctgc ctctgatcct gggtcctgtg ctggccttgc cggtcctggt
     1381 ggccctgggt gctctaggct tgtggcgtgt ccggcggagg caggagaagc agcgggattt
     1441 gcacagtgac ctgggcgagt ccagtctcat cctgaaggca tctgaacagg cagacagcat
     1501 gttgggggac ttcctggaca gcgactgtac tacgggcagc ggctcggggc tccccttctt
     1561 ggtgcagagg acggtagctc ggcaggttgc gctggtagag tgtgtgggaa agggccgata
     1621 tggtgaggtg tggcgcggtt cgtggcatgg cgagagtgtg gcggtcaaga ttttctcctc
     1681 acgagatgag cagtcctggt tccgggagac ggagatctac aacacagttc tgcttagaca
     1741 cgacaacatc ctaggcttca tcgcctccga catgacttcg cggaactcga gcacgcagct
     1801 gtggctcatc acccactacc atgaacacgg ctccctctat gactttctgc agaggcagac
     1861 gctggagccc cagttggccc tgaggctagc tgtgtccgcg gcctgcggcc tggcgcacct
     1921 acatgtggag atctttggca ctcaaggcaa accagccatt gcccatcgtg acctcaagag
     1981 tcgcaatgtg ctggtcaaga gtaacttgca gtgttgcatt gcagacctgg gactggctgt
     2041 gatgcactca caaagcagcg attacctgga tatcggcaac aacccccgag tgggtaccaa
     2101 aagatacatg gcacccgagg tgctggatga gcacatccgc acagactgct ttgagtcgta
     2161 caagtggaca gacatctggg cctttggcct agtgctatgg gagatcgccc ggcggaccat
     2221 catcaatggc attgtggagg attacaggcc acctttctat gacatggtac ccaatgaccc
     2281 cagttttgag gacatgaaaa aggtggtgtg cgttgaccag cagacaccca ccatccctaa
     2341 ccggctggct gcagatccgg tcctctccgg gctggcccag atgatgagag agtgctggta
     2401 ccccaacccc tctgctcgcc tcaccgcact gcgcataaag aagacattgc agaagctcag
     2461 tcacaatcca gagaagccca aagtgattca cctcgaggac tacaaagacc atgacggtga
     2521 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     2581 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2641 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2701 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2761 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2821 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2881 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2941 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3001 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3061 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3121 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3181 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3241 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3301 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3421 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3481 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3541 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3601 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3661 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3721 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3781 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3841 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3901 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3961 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4021 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4081 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4141 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4201 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4261 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4321 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4381 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4441 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4501 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4561 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4621 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4681 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4741 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4801 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4861 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4921 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4981 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5761 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5821 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5881 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5941 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6001 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6061 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6121 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6181 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6241 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6301 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6361 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6421 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6481 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6541 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6601 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6661 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6721 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6781 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6841 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6901 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6961 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
