LOCUS       62056_20450        9564 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9564)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1..1017
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             1072..2721
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     polyA_signal    2938..2993
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     promoter        complement(3696..3714)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3735..3751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3759..3775)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3783..3813)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3828..3849)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4137..4725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4899..5756)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5757..5861)
                     /label=AmpR promoter
     primer_bind     6335..6351
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6387..6886
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     intron          6954..7050
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             7117..8136
                     /codon_start=1
                     /label=SB100X
                     /note="hyperactive variant of the Sleeping Beauty
                     transposase (Mates et al., 2009)"
                     /translation="MGKSKEISQDLRKKIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTAAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGRSARKKPLLQNRHKKARLRFARAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMRKENYVDIL
                     KQHLKTSVRKLKLGRKWVFQMDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPTYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     polyA_signal    complement(8153..8287)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        8556..8574
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        9184..9563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          9564
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
ORIGIN
        1 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
       61 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      121 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      181 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      241 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      301 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
      361 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
      421 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
      481 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
      541 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
      601 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
      661 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
      721 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
      781 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
      841 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
      901 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      961 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1021 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt catggaagac
     1081 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tagaggatgg aaccgctgga
     1141 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
     1201 gatgcacata tcgaggtgaa catcacgtac gcggaatact tcgaaatgtc cgttcggttg
     1261 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     1321 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     1381 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgaacat ttcgcagcct
     1441 accgtagtgt ttgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaaatta
     1501 ccaataatcc agaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     1561 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtacca
     1621 gagtcctttg atcgtgacaa aacaattgca ctgataatga actcctctgg atctactggg
     1681 ttacctaagg gtgtggccct tccgcataga actgcctgcg tcagattctc gcatgccaga
     1741 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     1801 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     1861 ttaatgtata gatttgaaga agagctgttt ttacgatccc ttcaggatta caaaattcaa
     1921 agtgcgttgc tagtaccaac cctattttca ttcttcgcca aaagcactct gattgacaaa
     1981 tacgatttat ctaatttaca cgaaattgct tctgggggcg cacctctttc gaaagaagtc
     2041 ggggaagcgg ttgcaaaacg cttccatctt ccagggatac gacaaggata tgggctcact
     2101 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     2161 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     2221 gttaatcaga gaggcgaatt atgtgtcaga ggacctatga ttatgtccgg ttatgtaaac
     2281 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     2341 gcttactggg acgaagacga acacttcttc atagttgacc gcttgaagtc tttaattaaa
     2401 tacaaaggat accaggtggc ccccgctgaa ttggagtcga tattgttaca acaccccaac
     2461 atcttcgacg cgggcgtggc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     2521 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     2581 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     2641 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     2701 aagggcggaa agtccaaatt gtaaaatgta actgtattca gcgatgacga aattcttagc
     2761 tattgtaata ctctaggggg gatccactag aattcactcc tcaggtgcag gctgcctatc
     2821 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     2881 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2941 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     3001 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3061 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3121 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3181 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3241 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3301 ctcttctctt atggagatcc ctcgacctgc agcccaagct tgtggaaggc tactcgaaat
     3361 gtttgaccca agttaaacaa tttaaaggca atgctaccaa atactaattg agtgtatgta
     3421 aacttctgac ccactgggaa tgtgatgaaa gaaataaaag ctgaaatgaa tcattctctc
     3481 tactattatt ctgatatttc acattcttaa aataaagtgg tgatcctaac tgacctaaga
     3541 cagggaattt ttactaggat taaatgtcag gaattgtgaa aaagtgagtt taaatgtatt
     3601 tggctaaggt gtatgtaaac ttccgacttc aactgtatag ggatcctcta gctagagtcg
     3661 acctcgaggg ggggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     3721 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3781 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3841 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3901 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3961 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4021 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4201 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaaccgtg
     4321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4801 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4861 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4921 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4981 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5041 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5101 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5161 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5221 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5281 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5341 gttgtcagaa gtaacttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5401 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5461 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5521 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5581 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5641 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5701 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5761 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5821 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5881 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5941 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6001 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6061 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6121 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6181 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6241 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6301 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag
     6361 ctcggtaccc gggggcgcgc cctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     6421 agcatgcgct ttagcggccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     6481 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     6541 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     6601 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     6661 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     6721 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     6781 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     6841 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcatccatct
     6901 agatccttaa ttaagtctag agtcgaggaa ctgaaaaacc agaaagttaa ctggtaagtt
     6961 tagtcttttt gtcttttatt tcaggtcccg gatccggtgg tggtgcaaat caaagaactg
     7021 ctcctcagtg gatgttgcct ttacttctag gcctgtacgg aagtgttact tctgctctaa
     7081 aagctgcgga attgtacccg cggccgatcc gacatcatgg gaaaatcaaa agaaatcagc
     7141 caagacctca gaaaaaaaat tgtagacctc cacaagtctg gttcatcctt gggagcaatt
     7201 tccaaacgcc tggccgtacc acgttcatct gtacaaacaa tagtacgcaa gtataaacac
     7261 catgggacca cgcagccgtc ataccgctca ggaaggagac gcgttctgtc tcctagagat
     7321 gaacgtactt tggtgcgaaa agtgcaaatc aatcccagaa cagccgcaaa ggaccttgtg
     7381 aagatgctgg aggaaacagg tacaaaagta tctatatcca cagtaaaacg agtcctatat
     7441 cgacataacc tgaaaggccg ctcagcaagg aagaagccac tgctccaaaa ccgacataag
     7501 aaagccagac tacggtttgc aagagcacat ggggacaaag atcgtacttt ttggagaaat
     7561 gtcctctggt ctgatgaaac aaaaatagaa ctgtttggcc ataatgacca tcgttatgtt
     7621 tggaggaaga agggggaggc ttgcaagccg aagaacacca tcccaaccgt gaagcacggg
     7681 ggtggcagca tcatgttgtg ggggtgcttt gctgcaggag ggactggtgc acttcacaaa
     7741 atagatggca tcatgaggaa ggaaaattat gtggatatat tgaagcaaca tctcaagaca
     7801 tcagtcagga agttaaagct tggtcgcaaa tgggtcttcc aaatggacaa tgaccccaag
     7861 catacttcca aagttgtggc aaaatggctt aaggacaaca aagtcaaggt attggagtgg
     7921 ccatcacaaa gccctgacct caatcctata gaaaatttgt gggcagaact gaaaaagcgt
     7981 gtgcgagcaa ggaggcctac aaacctgact cagttacacc agctctgtca ggaggaatgg
     8041 gccaaaattc acccaactta ttgtgggaag cttgtggaag gctacccgaa acgtttgacc
     8101 caagttaaac aatttaaagg caatgctacc aaatactaga attggccgcg gggatccaga
     8161 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     8221 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     8281 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     8341 ggttttttcg gatcctctag agtcgactgt ttattaacgc ttacaatttc cattcgccat
     8401 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     8461 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     8521 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
     8581 tggagctcgg atccctatac agttgaagtc ggaagtttac atacacttaa gttggagtca
     8641 ttaaaactcg tttttcaact actccacaaa tttcttgtta acaaacaata gttttggcaa
     8701 gtcagttagg acatctactt tgtgcatgac acaagtcatt tttccaacaa ttgtttacag
     8761 acagattatt tcacttataa ttcactgtat cacaattcca gtgggtcaga agtttacata
     8821 cactaagttg actgtgcctt taaacagctt ggaaaattcc agaaaatgat gtcatggctt
     8881 tagaagctag atctggccat ctagagcggc cgcgcgcact agtgaattcc atggatatca
     8941 agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca
     9001 gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct
     9061 cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc
     9121 tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg tccctctaga
     9181 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     9241 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     9301 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     9361 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     9421 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     9481 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     9541 tattagtcat cgctattacc atgg
//
