LOCUS       V032253                 7012 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032253
VERSION     V032253
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..2502
                     /label="ACVR2A(NM_001278579)"
                     /note="ACVR2A(NM_001278579)"
                     /gene="ACVR2A"
     CDS             2509..2574
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2612..2836
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2882..3310
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3324..3653
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3720..4514
                     /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    4688..4809
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4858..4874)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4882..4898
                     /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(4906..4936)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4951..4972
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5260..5845)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6016..6876)
                     /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(6877..6981)
                     /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 accatgggag ctgctgcaaa gttggcgttt gccgtctttc ttatctcctg ttcttcaggt
     1021 gctatacttg gtagatcaga aactcaggag tgtcttttct ttaatgctaa ttgggaaaaa
     1081 gacagaacca atcaaactgg tgttgaaccg tgttatggtg acaaagataa acggcggcat
     1141 tgttttgcta cctggaagaa tatttctggt tccattgaaa tagtgaaaca aggttgttgg
     1201 ctggatgata tcaactgcta tgacaggact gattgtgtag aaaaaaaaga cagccctgaa
     1261 gtatattttt gttgctgtga gggcaatatg tgtaatgaaa agttttctta ttttccggag
     1321 atggaagtca cacagcccac ttcaaatcca gttacaccta agccacccta ttacaacatc
     1381 ctgctctatt ccttggtgcc acttatgtta attgcgggga ttgtcatttg tgcattttgg
     1441 gtgtacaggc atcacaagat ggcctaccct cctgtacttg ttccaactca agacccagga
     1501 ccacccccac cttctccatt actaggtttg aaaccactgc agttattaga agtgaaagca
     1561 aggggaagat ttggttgtgt ctggaaagcc cagttgctta acgaatatgt ggctgtcaaa
     1621 atatttccaa tacaggacaa acagtcatgg caaaatgaat acgaagtcta cagtttgcct
     1681 ggaatgaagc atgagaacat attacagttc attggtgcag aaaaacgagg caccagtgtt
     1741 gatgtggatc tttggctgat cacagcattt catgaaaagg gttcactatc agactttctt
     1801 aaggctaatg tggtctcttg gaatgaactg tgtcatattg cagaaaccat ggctagagga
     1861 ttggcatatt tacatgagga tatacctggc ctaaaagatg gccacaaacc tgccatatct
     1921 cacagggaca tcaaaagtaa aaatgtgctg ttgaaaaaca acctgacagc ttgcattgct
     1981 gactttgggt tggccttaaa atttgaggct ggcaagtctg caggcgatac ccatggacag
     2041 gttggtaccc ggaggtacat ggctccagag gtattagagg gtgctataaa cttccaaagg
     2101 gatgcatttt tgaggataga tatgtatgcc atgggattag tcctatggga actggcttct
     2161 cgctgtactg ctgcagatgg acctgtagat gaatacatgt tgccatttga ggaggaaatt
     2221 ggccagcatc catctcttga agacatgcag gaagttgttg tgcataaaaa aaagaggcct
     2281 gttttaagag attattggca gaaacatgct ggaatggcaa tgctctgtga aaccattgaa
     2341 gaatgttggg atcacgacgc agaagccagg ttatcagctg gatgtgtagg tgaaagaatt
     2401 acccagatgc agagactaac aaatattatt accacagagg acattgtaac agtggtcaca
     2461 atggtgacaa atgttgactt tcctcccaaa gaatctagtc tactcgagga ctacaaagac
     2521 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga
     2581 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2641 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2701 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2761 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2821 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2881 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2941 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3001 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3061 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3121 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3181 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3241 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3301 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3481 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3541 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3601 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3661 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3721 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3781 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3841 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3901 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3961 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4021 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4081 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4141 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4201 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4261 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4321 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4381 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4441 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4501 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4561 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4621 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4681 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4741 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4801 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5701 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     5821 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5881 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5941 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6001 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6061 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
