LOCUS       V025680                 8054 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025680
VERSION     V025680
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3550
                     /label="BRAT1(NM_001350626)"
                     /note="BRAT1(NM_001350626)"
                     /gene="BRAT1"
     CDS             3557..3574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3621..3845
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3891..4319
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4333..4662
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4729..5523
                     /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    5697..5818
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5867..5883)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5891..5907
                     /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(5915..5945)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5960..5981
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6269..6854)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7025..7885)
                     /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(7886..7990)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gacccagaat gcgcccagct gctcccggct ctctgtgctg ttctggtaga
      961 tcccaggcag ccggtggcag atgacacctg tttggagaag ctcctggact ggtttaaaac
     1021 ggtcactgaa ggagagtcca gtgtcgtgct gctgcaggag cacccctgcc tggtggagct
     1081 gctgtcccat gtgctgaaag tccaggacct gagttctggg gtcctctcct tctcactgcg
     1141 cctggcagga accttcgcag cccaggaaaa ctgcttccag tatcttcagc agggggagtt
     1201 actaccaggg ctctttgggg agccaggacc cctcggccga gcaacctggg ccgtccccac
     1261 cgtgcgcagc ggctggatcc agggcctgcg ctccctggca cagcacccca gcgccctgcg
     1321 cttcctggcc gaccatggtg cggtcgacac catcttctcc ctgcagggag actccagcct
     1381 gtttgtggcc tcggcggcca gtcagctcct ggtgcacgtc ctggctttgt ccatgcgagg
     1441 tggagccgag gggcagccct gcctgccggg gggtgactgg cccgcgtgtg cccagaagat
     1501 catggatcac gttgaagagt ccttgtgctc cgcggccacc cccaaggtca ctcaggccct
     1561 gaacgtcctg accacgacct tcgggcgctg ccagagcccc tggacggaag ccctgtgggt
     1621 gcggctgagt ccccgcgtgg cctgtctgct ggagagagac cccatccccg ccgcacactc
     1681 gttcgtggac ctgcttctct gtgtggctcg ttctcccgtg ttcagttctt ccgacggcag
     1741 cctgtgggag acagtggcgc gggctctgag ctgcctgggt cccacccaca tgggacccct
     1801 ggctttgggg atcctgaagc tcgagcactg tccacaggca ctgaggaccc aggccttcca
     1861 ggtccttctc cagcccctgg cctgtgtcct gaaggccacg gttcaggccc ccggaccccc
     1921 aggcttgctg gacgggacgg cagacgatgc cacgacggtg gacacactcc tggcctccaa
     1981 gtcgtcctgc gccggcctcc tgtgccgcac cctggctcac ctggaggagc tgcagccgct
     2041 gccccagcgc ccttcaccgt ggccccaggc gtctctactg ggggctacag tgactgtcct
     2101 gcggctctgt gacggctcgg ctgcccctgc ctccagtgtg gggggccacc tctgtgggac
     2161 cctggcgggc tgcgtccggg tccagcgagc agccctcgac ttcctgggga cgctgtcaca
     2221 ggggacaggc ccccaggagc tggtgacgca ggcgcttgct gtcctcctgg agtgcctcga
     2281 gagccccggc tccagcccca cggttctgaa gaaggccttc caggccacgc tcaggtggct
     2341 cctgagctca cccaagaccc ccggctgctc tgatctcggc cccctcatcc cgcagttcct
     2401 cagagagctg ttccctgtgc tgcagaaacg cctgtgccac ccctgctggg aggtgaggga
     2461 ctccgccctc gagttcctga cccagctgag caggcactgg ggaggacagg ctgacttcag
     2521 atgcgcactc ttggcttcag aggtgcctca gctggccctg cagctcctcc aggaccctga
     2581 gagttatgtc cgagcgagtg cagtgaccgc catggggcag ctgtccagcc agggcctgca
     2641 cgcccccacc agccctgagc atgcagaggc ccggcagatg ggtaaactga ggattggagg
     2701 gccgtgtgct cactgcgcag catgggaagg tgtcagagcg ggctgtggcc ctagactgca
     2761 tgtccggggc cagcccccat catgtaccgg agtcctgttg cgggagcctc gctcctgcca
     2821 ccccaccaac catccgcact tgctccctgt cccccagagc ctgttcctgg agctcctgca
     2881 catcctctcc gtagactcgg agggcttccc acggcgggcg gtcatgcaag tcttcactga
     2941 gtggctgcgg gacggccacg ccgacgcggc ccaggacacg gagcagttcg tggccactgt
     3001 gctgcaggcg gcgagccgag acctggactg ggaggtccgc gcccagggcc tggagctggc
     3061 cctcgtgttc ctgggccaga ctttggggcc gccgcgtacc cactgcccct atgccgtggc
     3121 cctacccgag gtggccccag cccagccact caccgaggca ctgagggctc tctgccacgt
     3181 ggggctcttt gacttcgcct tttgtgcctt gtttgactgc gaccgccctg tggcgcagaa
     3241 gtcttgtgac ctccttctct tcctgaggga caagattgct tcctacagca gcctgcggga
     3301 ggccaggggc agccccaaca ctgcctccgc agaggccacc ctgccgaggt ggcgggcggg
     3361 tgagcaggcc cagcccccag gggaccagga gcctgaggct gtgctggcca tgctcaggtc
     3421 cctagacctg gagggcctgc ggagcacgct ggccgagagc agcgaccacg tggaaaagag
     3481 tccccagtcc ctcctgcagg acatgctggc cacgggaggc ttcctgcagg gggacgaggc
     3541 cgactgctac gaattccatc accatcatca ccattgactc gagtctagag ggcccgttta
     3601 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3661 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3721 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3781 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3841 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3901 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3961 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4021 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4081 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4141 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4201 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4261 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4321 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4381 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4501 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4561 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4621 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4681 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4741 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4801 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4861 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4921 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4981 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5041 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5101 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5161 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5221 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5281 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5341 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5401 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5461 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5521 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5581 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5641 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5701 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5761 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5821 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5881 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5941 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6001 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6061 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6121 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6181 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6241 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6301 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6361 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6421 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6481 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6541 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6601 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6661 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6721 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6781 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6841 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6901 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6961 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7021 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7081 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7141 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7201 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7261 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7321 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7381 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7441 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7501 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7561 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7621 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7681 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7741 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7801 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7861 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7921 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7981 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8041 cgatccccta tggt
//
