LOCUS       V035329                 6848 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035329
VERSION     V035329
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             955..2748
                     /label="NR4A2(NM_006186)"
                     /note="NR4A2(NM_006186)"
                     /gene="NR4A2"
     CDS             2770..2799
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2815..2832
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2861..3085
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3131..3559
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3573..3903
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        3951..3998
                     /note="synthetic bacterial promoter&nbsp;"
                     /label="EM7 promoter"
     CDS             4017..4391
                     /codon_start=1
                     /gene="Sh&nbsp;ble from&nbsp;Streptoalloteichus
                     hindustanus"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin™"
                     /product="antibiotic-binding protein"
                     /transl_table=1
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD*"
                     /label="BleoR"
     polyA_signal    4521..4642
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4691..4707)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4715..4731
                     /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(4739..4769)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4784..4805
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5093..5681)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5852..6712)
                     /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(6713..6817)
                     /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 ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatgcct
      961 tgtgttcagg cgcagtatgg gtcctcgcct caaggagcca gccccgcttc tcagagctac
     1021 agttaccact cttcgggaga atacagctcc gatttcttaa ctccagagtt tgtcaagttt
     1081 agcatggacc tcaccaacac tgaaatcact gccaccactt ctctccccag cttcagtacc
     1141 tttatggaca actacagcac aggctacgac gtcaagccac cttgcttgta ccaaatgccc
     1201 ctgtccggac agcagtcctc cattaaggta gaagacattc agatgcacaa ctaccagcaa
     1261 cacagccacc tgccccccca gtctgaggag atgatgccgc actccgggtc ggtttactac
     1321 aagccctcct cgcccccgac gcccaccacc ccgggcttcc aggtgcagca cagccccatg
     1381 tgggacgacc cgggatctct ccacaacttc caccagaact acgtggccac tacgcacatg
     1441 atcgagcaga ggaaaacgcc agtctcccgc ctctccctct tctcctttaa gcaatcgccc
     1501 cctggcaccc cggtgtctag ttgccagatg cgcttcgacg ggcccctgca cgtccccatg
     1561 aacccggagc ccgccggcag ccaccacgtg gtggacgggc agaccttcgc tgtgcccaac
     1621 cccattcgca agcccgcgtc catgggcttc ccgggcctgc agatcggcca cgcgtctcag
     1681 ctgctcgaca cgcaggtgcc ctcaccgccg tcgcggggct ccccctccaa cgaggggctg
     1741 tgcgctgtgt gtggggacaa cgcggcctgc caacactacg gcgtgcgcac ctgtgagggc
     1801 tgcaaaggct tctttaagcg cacagtgcaa aaaaatgcaa aatacgtgtg tttagcaaat
     1861 aaaaactgcc cagtggacaa gcgtcgccgg aatcgctgtc agtactgccg atttcagaag
     1921 tgcctggctg ttgggatggt caaagaagtg gttcgcacag acagtttaaa aggccggaga
     1981 ggtcgtttgc cctcgaaacc gaagagccca caggagccct ctcccccttc gcccccggtg
     2041 agtctgatca gtgccctcgt cagggcccat gtcgactcca acccggctat gaccagcctg
     2101 gactattcca ggttccaggc gaaccctgac tatcaaatga gtggagatga cacccagcat
     2161 atccagcaat tctatgatct cctgactggc tccatggaga tcatccgggg ctgggcagag
     2221 aagatccctg gcttcgcaga cctgcccaaa gccgaccaag acctgctttt tgaatcagct
     2281 ttcttagaac tgtttgtcct tcgattagca tacaggtcca acccagtgga gggtaaactc
     2341 atcttttgca atggggtggt cttgcacagg ttgcaatgcg ttcgtggctt tggggaatgg
     2401 attgattcca ttgttgaatt ctcctccaac ttgcagaata tgaacatcga catttctgcc
     2461 ttctcctgca ttgctgccct ggctatggtc acagagagac acgggctcaa ggaacccaag
     2521 agagtggaag aactgcaaaa caagattgta aattgtctca aagaccacgt gactttcaac
     2581 aatggggggt tgaaccgccc caattatttg tccaaactgt tggggaagct cccagaactt
     2641 cgtacccttt gcacacaggg gctacagcgc attttctacc tgaaattgga agacttggtg
     2701 ccaccgccag caataattga caaacttttc ctggacactt tacctttcct cgagtctaga
     2761 gggcccttcg aacaaaaact catctcagaa gaggatctga atatgcatac cggtcatcat
     2821 caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2881 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2941 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3001 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3061 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3121 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3181 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3241 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     3301 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3361 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3421 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3481 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     3541 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3601 caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3721 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3781 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3841 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3901 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     3961 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     4021 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     4081 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     4141 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     4201 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     4261 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     4321 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     4381 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4441 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4501 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4621 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4681 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4741 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4801 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4861 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4921 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4981 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5041 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5101 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5161 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5221 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5281 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5341 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5401 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5461 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5521 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5581 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5641 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5701 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5761 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5821 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5881 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5941 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6001 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6061 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6121 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6181 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6241 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6301 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6361 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6421 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6481 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6541 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6601 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6661 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6721 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6781 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6841 ctgacgtc
//
