LOCUS       62056_9415        9920 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 9920)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             4607..4672
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4696..5284
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    5350..5398
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5600..6028
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6042..6371
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6438..7229
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    7408..7541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(7626..7656)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7671..7692)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7980..8568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8742..9599)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9600..9704)
                     /label=AmpR promoter
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgaccgc tcgcggcctg gcccttggcc tcctcctgct
      721 gctactgtgt ccagcgcagg tgttttcaca gtcctgtgtt tggtatggag agtgtggaat
      781 tgcatatggg gacaagaggt acaattgcga atattctggc ccaccaaaac cattgccaaa
      841 ggatggatat gacttagtgc aggaactctg tccaggattc ttctttggca atgtcagtct
      901 ctgttgtgat gttcggcagc ttcagacact aaaagacaac ctgcagctgc ctctacagtt
      961 tctgtccaga tgtccatcct gtttttataa cctactgaac ctgttttgtg agctgacatg
     1021 tagccctcga cagagtcagt ttttgaatgt tacagctact gaagattatg ttgatcctgt
     1081 tacaaaccag acgaaaacaa atgtgaaaga gttacaatac tacgtcggac agagttttgc
     1141 caatgcaatg tacaatgcct gccgggatgt ggaggccccc tcaagtaatg acaaggccct
     1201 gggactcctg tgtgggaagg acgctgacgc ctgtaatgcc accaactgga ttgaatacat
     1261 gttcaataag gacaatggac aggcaccttt taccatcact cctgtgtttt cagattttcc
     1321 agtccatggg atggagccca tgaacaatgc caccaaaggc tgtgacgagt ctgtggatga
     1381 ggtcacagca ccatgtagct gccaagactg ctctattgtc tgtggcccca agccccagcc
     1441 cccacctcct cctgctccct ggacgatcct tggcttggac gccatgtatg tcatcatgtg
     1501 gatcacctac atggcgtttt tgcttgtgtt ttttggagca ttttttgcag tgtggtgcta
     1561 cagaaaacgg tattttgtct ccgagtacac tcccatcgat agcaatatag ctttttctgt
     1621 taatgcaagt gacaaaggaa cggcttggct cttaacctcc accttccctt cctctcccgt
     1681 tcttccagga gaggcgtcct gctgtgaccc tgtcagcgca gcatttgagg gctgcttgag
     1741 gcggctgttc acacgctggg ggtctttctg cgtccgaaac cctggctgtg tcattttctt
     1801 ctcgctggtc ttcattactg cgtgttcgtc aggcctggtg tttgtccggg tcacaaccaa
     1861 tccagttgac ctctggtcag cccccagcag ccaggctcgc ctggaaaaag agtactttga
     1921 ccagcacttt gggcctttct tccggacgga gcagctcatc atccgggccc ctctcactga
     1981 caaacacatt taccagccat acccttcggg agctgatgta ccctttggac ctccgcttga
     2041 catacagata ctgcaccagg ttcttgactt acaaatagcc atcgaaaaca ttactgcctc
     2101 ttatgacaat gagactgtga cacttcaaga catctgcttg gcccctcttt caccgtataa
     2161 cacgaactgc accattttga gtgtgttaaa ttacttccag aacagccatt ccgtgctgga
     2221 ccacaagaaa ggggacgact tctttgtgta tgccgattac cacacgcact ttctgtactg
     2281 cgtacgggct cctgcctctc tgaatgatac aagtttgctc catgaccctt gtctgggtac
     2341 gtttggtgga ccagtgttcc cgtggcttgt gttgggaggc tatgatgatc aaaactacaa
     2401 taacgccact gcccttgtga ttaccttccc tgtcaataat tactataatg atacagagaa
     2461 gctccagagg gcccaggcct gggaaaaaga gtttattaat tttgtgaaaa actacaagaa
     2521 tcccaatctg accatttcct tcactgctga acgaagtatt gaagatgaac taaatcgtga
     2581 aagtgacagt gatgtcttca ccgttgtaat tagctatgcc atcatgtttc tatatatttc
     2641 cctagccttg gggcacatga aaagctgtcg caggcttctg gtggattcga aggtctcact
     2701 aggcatcgcg ggcatcttga tcgtgctgag ctcggtggct tgctccttgg gtgtcttcag
     2761 ctacattggg ttgcccttga ccctcattgt gattgaagtc atcccgttcc tggtgctggc
     2821 tgttggagtg gacaacatct tcattctggt gcaggcctac cagagagatg aacgtcttca
     2881 aggggaaacc ctggatcagc agctgggcag ggtcctagga gaagtggctc ccagtatgtt
     2941 cctgtcatcc ttttctgaga ctgtagcatt tttcttagga gcattgtccg tgatgccagc
     3001 cgtgcacacc ttctctctct ttgcgggatt ggcagtcttc attgactttc ttctgcagat
     3061 tacctgtttc gtgagtctct tggggttaga cattaaacgt caagagaaaa atcggctaga
     3121 catcttttgc tgtgtcagag gtgctgaaga tggaacaagc gtccaggcct cagagagctg
     3181 tttgtttcgc ttcttcaaaa actcctattc tccacttctg ctaaaggact ggatgagacc
     3241 aattgtgata gcaatatttg tgggtgttct gtcattcagc atcgcagtcc tgaacaaagt
     3301 agatattgga ttggatcagt ctctttcgat gccagatgac tcctacatgg tggattattt
     3361 caaatccatc agtcagtacc tgcatgcggg tccgcctgtg tactttgtcc tggaggaagg
     3421 gcacgactac acttcttcca aggggcagaa catggtgtgc ggcggcatgg gctgcaacaa
     3481 tgattccctg gtgcagcaga tatttaacgc ggcgcagctg gacaactata cccgaatagg
     3541 cttcgccccc tcgtcctgga tcgacgatta tttcgactgg gtgaagccac agtcgtcttg
     3601 ctgtcgagtg gacaatatca ctgaccagtt ctgcaatgct tcagtggttg accctgcctg
     3661 cgttcgctgc aggcctctga ctccggaagg caaacagagg cctcaggggg gagacttcat
     3721 gagattcctg cccatgttcc tttcggataa ccctaacccc aagtgtggca aagggggaca
     3781 tgctgcctat agttctgcag ttaacatcct ccttggccat ggcaccaggg tcggagccac
     3841 gtacttcatg acctaccaca ccgtgctgca gacctctgct gactttattg acgctctgaa
     3901 gaaagcccga cttatagcca gtaatgtcac cgaaaccatg ggcattaacg gcagtgccta
     3961 ccgagtattt ccttacagtg tgttttatgt cttctacgaa cagtacctga ccatcattga
     4021 cgacactatc ttcaacctcg gtgtgtccct gggcgcgata tttctggtga ccatggtcct
     4081 cctgggctgt gagctctggt ctgcagtcat catgtgtgcc accatcgcca tggtcttggt
     4141 caacatgttt ggagttatgt ggctctgggg catcagtctg aacgctgtat ccttggtcaa
     4201 cctggtgatg agctgtggca tctccgtgga gttctgcagc cacataacca gagcgttcac
     4261 ggtgagcatg aaaggcagcc gcgtggagcg cgcggaagag gcacttgccc acatgggcag
     4321 ctccgtgttc agtggaatca cacttacaaa atttggaggg attgtggtgt tggcttttgc
     4381 caaatctcaa attttccaga tattctactt caggatgtat ttggccatgg tcttactggg
     4441 agccactcac ggattaatat ttctccctgt cttactcagt tacatagggc catcagtaaa
     4501 taaagccaaa agttgtgcca ctgaagagcg atacaaagga acagagcgcg aacggcttct
     4561 aaatttcgaa ttctgcagat atccagcaca gtggcggccg ctcgaggact acaaagacca
     4621 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     4681 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4741 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4801 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4861 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4921 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4981 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5041 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     5101 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5161 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5221 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5281 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5341 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     5401 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5461 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5521 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     5581 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5641 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5701 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     5761 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5821 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5881 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5941 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     6001 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     6061 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6121 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6181 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6241 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6301 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6361 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     6421 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6481 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6541 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6601 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6661 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6721 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6781 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6841 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6901 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6961 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     7021 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     7081 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     7141 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     7201 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     7261 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     7321 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     7381 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     7441 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7501 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     7561 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7621 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7681 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7741 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7801 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7861 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7921 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7981 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     8041 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     8101 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     8161 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     8221 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8281 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8341 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8401 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8461 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8521 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8581 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8641 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8701 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8761 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8821 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8881 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8941 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     9001 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     9061 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     9121 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     9181 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     9241 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     9301 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     9361 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     9421 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9481 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9541 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9601 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9661 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9721 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     9781 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     9841 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     9901 gcatgaagaa tctgcttagg
//
