LOCUS       V041133                 7942 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041133
VERSION     V041133
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             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..3432
                     /label="Nr4a1(NM_024388)"
                     /note="Nr4a1(NM_024388)"
                     /gene="Nr4a1"
     polyA_signal    3509..3733
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3779..4207
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4221..4550
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4617..5411
                     /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    5585..5706
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5755..5771)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5779..5795
                     /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(5803..5833)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5848..5869
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6157..6742)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6913..7773)
                     /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(7774..7878)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgccctgt atccaagctc aatatggaac accagcaacc
     1681 agcccaggac cgcgtgacca cctgacaggt gaccccctgg cccttgagtt cagcaagccc
     1741 accatggacc tggccagccc cgagacggcg cccaccgcac ctgccaccct gcccagtttc
     1801 agcaccttca tggacggcgg atacaccgga gagtttgaca ccttcctata ccagctgccg
     1861 gggacagccc agccgtgctc ctcggcctcc tccacatctt cttcctcgtc ctcggccacc
     1921 tctcccgcgt cggcttcctt taagtttgag gactttcagg tgtatggctg ctaccctggc
     1981 accctgagcg gcccattaga cgagaccctg tcctccagtg gctctgatta ctatggaagc
     2041 ccctgctcag ccccgtcacc acctacaccc aacttccagc catcccagct ctccccttgg
     2101 gatggttcat ttggccactt ctcccccagc cagacttatg aaggccttcg ggtatggact
     2161 gagcagttgc ccaaggcttc cgggcctccg ccacctccaa ccttcttctc cttcagtcct
     2221 cccaccggtc ccagccccag cctggcccag agttccctga agttgttccc agcaccagcc
     2281 acccaccagc taggggaggg ggagagctat tccgtgccag ccgctttccc gggcttggca
     2341 cccacctctc cgaactgtga cacttccggc attctggacg cacccgtaac ctccaccaag
     2401 gcccgcagtg ggtcttcagg gggcagtgag ggccgctgtg ctgtctgtgg agacaacgct
     2461 tcgtgccagc attatggggt ccgcacctgt gagggctgca aaggcttctt caagcgcaca
     2521 gtacagaaaa gtgccaagta catctgcctg gcaaacaagg actgccctgt ggacaagagg
     2581 cggcggaacc gctgccagtt ctgtcgcttc cagaagtgcc tggctgtggg catggtgaag
     2641 gaagttgtcc ggacagacag cctaaagggg cggcggggcc ggctaccttc aaaacccaag
     2701 cagcccccag atgcctcccc taccaatctc ctcacgtccc tcattcgagc acatttggac
     2761 tccgggccta acactgccaa gttggactat tccaagttcc aggaactggt gctgccccgc
     2821 tttggaaagg aagatgccgg tgacgtgcag caattttatg acttgctctc tggctccctg
     2881 gacgtcatcc ggaagtgggc ggaaaagatc ccgggcttca ttgagctttc cccaggagac
     2941 caagacctgt tgctagagtc cgcctttctg gagctcttta tcctccgcct ggcctaccga
     3001 tctaagcccg gtgaggggaa gctcatcttc tgctcaggcc tggtgctaca ccggctgcag
     3061 tgtgcccgcg gctttggtga ctggatagac aacattctag ccttctcacg gtccctgcac
     3121 agcttgggtg ttgatgttcc tgcctttgcc tgcctgtctg ctctggtcct catcactgat
     3181 cgacatggcc tccaggaccc tcggcgggtg gaggagctgc agaaccgcat tgctagctgt
     3241 ctgaaggaac acatggccgc cgtggcagga gacccacagc cggccagctg cctgtcacgt
     3301 ctgctgggca aactgcctga gctccggact ctgtgcaccc aaggcctaca gcgcatcttt
     3361 tgcctcaagc tggaggactt ggtgccccct ccacctatcg tggacaagat ctttatggac
     3421 accttgtctt tctgaggatc ctgatatcca gcacagtggc ggccgctcga gtctagaggg
     3481 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3541 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3601 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3661 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3721 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3781 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3841 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3901 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3961 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4021 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4081 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4141 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4201 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4381 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4441 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4501 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4561 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4621 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4681 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4741 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4801 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4861 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4921 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4981 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5041 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5101 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5161 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5221 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5281 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5341 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5401 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5461 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5521 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5581 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5641 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5701 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5761 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5821 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5881 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5941 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6001 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6061 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6121 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6421 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6481 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6541 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6601 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6661 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     6721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7921 agatctcccg atcccctatg gt
//
