LOCUS       62056_6905        8622 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 8622)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             682..705
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             721..2214
                     /codon_start=1
                     /label=CRY2PHR
                     /note="N-terminal photolyase homology region of Arabidopsis
                     thaliana cryptochrome 2"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     primer_bind     complement(2967..2983)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3040..3058)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3332..3453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3472..4650
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             4688..5404
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    5494..5542
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      complement(5544..5999)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6026..6128
                     /label=AmpR promoter
     promoter        6132..6489
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6524..7315
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      7926..8514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccaccat ggattacaag gatgacgacg ataaggcccg ggcggatccc
      721 atgaagatgg acaagaaaac aatcgtgtgg tttagaagag atttgaggat tgaggacaat
      781 cccgctctgg ctgctgccgc tcatgaagga agtgtgtttc cagtgttcat ctggtgcccc
      841 gaggaggaag gacagttcta ccctggacgg gcttccagat ggtggatgaa gcagagcctg
      901 gcccatctgt ctcagtccct gaaggctctg gggtctgacc tgactctgat taagacccac
      961 aacaccatca gcgctatcct ggattgtatt cgcgtgaccg gcgccacaaa ggtggtgttc
     1021 aaccacctgt acgaccccgt gtccctggtg agagatcaca cagtgaagga aaaactggtg
     1081 gagagaggaa tcagcgtgca gtcctacaac ggcgacctgc tgtacgagcc ttgggaaatc
     1141 tactgcgaga aaggcaagcc cttcaccagc tttaacagct attggaagaa gtgcctggac
     1201 atgagcatcg agagcgtgat gctgccaccc ccttggagac tgatgcctat taccgccgcc
     1261 gctgaggcta tctgggcttg ttctatcgag gagctgggcc tggagaatga ggctgaaaag
     1321 cccagcaacg ccctgctgac tagagcttgg agccctggat ggagcaacgc tgataagctg
     1381 ctgaacgagt ttattgagaa gcagctgatc gactatgcca agaacagcaa gaaggtggtg
     1441 ggaaactcca ccagcctgct gagtccttac ctgcacttcg gagaaattag cgtgcggcac
     1501 gtgtttcagt gcgccagaat gaaacagatt atctgggcca gagacaaaaa cagcgaaggc
     1561 gaagagagcg ccgacctgtt tctgagaggc atcggactga gggagtactc taggtacatt
     1621 tgcttcaact tccccttcac ccacgagcag agcctgctga gccatctgag attcttcccc
     1681 tgggacgccg acgtggataa gtttaaggcc tggagacagg gccggaccgg ataccctctg
     1741 gtggatgctg gaatgagaga gctgtgggcc accggatgga tgcataacag gattagagtg
     1801 attgtgagca gcttcgccgt gaagttcctg ctgctgcctt ggaagtgggg catgaaatat
     1861 ttctgggaca ccctgctgga cgctgacctg gaatgtgaca tcctgggctg gcagtatatc
     1921 tctggatcaa tccctgatgg acacgaactg gacagactgg acaaccccgc tctgcaagga
     1981 gctaaatacg accctgaggg agagtacatc cggcagtggc tgcctgaact ggctagactg
     2041 cctaccgagt ggattcatca cccctgggac gctcctctga ccgtgctgaa ggcttctgga
     2101 gtggagctgg gaaccaacta tgccaagccc atcgtggaca tcgacaccgc tagagaactg
     2161 ctggccaagg ctatctctcg caccagagaa gcccagatca tgatcggagc cgctggagga
     2221 ggaggatctg gaggaggagg ctctggagga ggcggatctg ataaaacaca cacatgcccc
     2281 ccatgccctg ctcctgaact gctgggagga ccttctgtgt tcctgtttcc tcccaagcct
     2341 aaggacaccc tgatgatcag ccggacccct gaagtgacct gcgtggtggt ggatgtgagc
     2401 catgaagatc ccgaagtgaa gtttaactgg tacgtggacg gcgtggaagt gcacaacgct
     2461 aagaccaaac ccagagagga acagtacaac agtacataca gagtggtgag cgtgctgaca
     2521 gtgctgcacc aggattggct gaacggcaag gagtacaagt gcaaggtgag caacaaggcc
     2581 ctgcccgctc ctattgagaa gaccatcagc aaagccaagg gacagcccag ggagcctcag
     2641 gtgtatacac tgcctcccag cagagaagag atgaccaaga accaggtgag cctgacctgc
     2701 ctggtgaaag gattctaccc cagcgacatc gccgtggaat gggaatctaa tggccagccc
     2761 gagaacaact acaaaacaac cccacccgtg ctggactcag acggatcttt cttcctgtac
     2821 agcaaactga ccgtggacaa gagcaggtgg cagcagggaa acgtgttcag ctgctccgtg
     2881 atgcacgagg ctctgcataa ccactatacc cagaagagcc tgagcctgtc ccccggaaaa
     2941 tgatgagaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc
     3001 ttaattaatt aaggtaccag gtaagtgtac ccaattcgcc ctatagtgag tcgtattaca
     3061 attcactcga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg agatccaatt
     3121 tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt agattcacag
     3181 tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca
     3241 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3301 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3361 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3421 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta ggcctactag tggctccggt
     3481 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     3541 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     3601 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     3661 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     3721 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
     3781 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     3841 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     3901 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     3961 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     4021 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     4081 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     4141 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     4201 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     4261 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     4321 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     4381 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     4441 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     4501 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     4561 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     4621 agtttttttc ttccatttca ggtgtcgtga ctctagagct agcgtcgaca cgcgtgcggc
     4681 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4741 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     4801 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     4861 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     4921 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     4981 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     5041 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     5101 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     5161 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     5221 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     5281 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     5341 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     5401 caagtaaggt accaccggtt agtaatgagt ttaaacgggg gaggctaact gaaacacgga
     5461 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     5521 acgggtgttg ggtcgtttgt tcaaaattgt aagcgttaat attttgttaa aattcgcgtt
     5581 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5641 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5701 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5761 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5821 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5881 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5941 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6001 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6121 aaggaagaat cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     6181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6241 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6301 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6361 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6421 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6481 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     6541 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6601 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6661 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     6721 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6781 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6841 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6901 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6961 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagaacatc aggggctcgc
     7021 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     7081 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7141 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     7201 tgatattgct gaagaacttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     7261 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     7321 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     7381 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     7441 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     7501 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     7561 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     7621 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     7681 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     7741 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     7801 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7861 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7921 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7981 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8041 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     8101 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8161 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8221 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8281 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8341 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8401 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8461 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8521 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8581 cctgcgttat cccctgattc tgtggataac cgtattaccg cc
//
