LOCUS       62056_9265        7628 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 7628)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7628
                     /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             2315..2380
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2404..2992
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3058..3106
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3308..3736
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3750..4079
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4146..4937
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5116..5249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5286..5302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5310..5326)
                     /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(5334..5364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5379..5400)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5688..6276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6450..7307)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7308..7412)
                     /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 ccatggagag agagaagaac gtggtgtacg gccccgagcc
      721 caagcaccct ctgggcaact tcaccgccgg cgagatgctg tacaacgctc tgcacaagca
      781 ctcccacatc ccccaggcca tcctggacgt gatgggcaac gagtcccttt cctaccagga
      841 gttcttcgac actactgtga agctgggcca gagcctccag aactgtggct acaagatgaa
      901 cgatgtcgtg tcgatctgtg cagagaacaa caagagattc ttcatcccca tcatctccgc
      961 ctggtacatc ggcatggtgg tggcccctgt gaacgaggac tatatcccag acgagctgtg
     1021 taaagtgacc ggcatctcca agccgatcct ggtcttcacc actaggaaga tcctgcctaa
     1081 ggttttggag gttaaagaca gaaccaacta cataaagagg atcatcatac tggactctga
     1141 agagaacctg ctgggctgcg agagcctgca caacttcatg tccaggtact ccgacaacaa
     1201 cctccaaaca ttcaagcctc tgcactacga ccctgtggac caggtagccg ccatcctgtg
     1261 ctcctccggc acaaccggcc tgcctaaagg cgtgatgcag acccacagga acatctgtgt
     1321 gagactcaca cacgcatctg accccagagt gggtacacaa ctcatccccg gcgtatccgt
     1381 gctggcctac ctgccattct tccacgcctt cggcttcagt atcaacctgg gctatttcat
     1441 ggtgggcctg agagtggtga tgctccgaag gtttaaccag gaggtgttcc tgaaggccat
     1501 ccaggactac gaggtgagga gcgtgatcaa cgttccctcc acaatcctgt tcctgtccaa
     1561 gagccctctg gtggacaagt acgacctatc caccctggcg gagctgtgct gtggagccgc
     1621 tcctctggcg aaggaggtgg ccgagatcgc cgtgaagagg ctgaacctgc cagggatacg
     1681 gtgtggctac ggtctaacag agtctacctc cgccaacatc catactctgc acaacgagtt
     1741 caagtccggc tccctgggca aggtgacacc ttacatggcc gccaagatca tcgacaggaa
     1801 caccggcgag gccctgggtc caaaccaggt gggcgagctg tgcatctggg gacctatggt
     1861 aacaaaaggc tatgtgaaca acccacaggc tactaaggag gccatcgacg acgacggctg
     1921 gctgcactct ggcgacttcg gctactacga cgaggacgag tatttctaca tcgtggaccg
     1981 gtacaaggag ctgatcaaat acaagggcta tcaggtcgcc cctgtggagc tggaggagat
     2041 cctccttcag cacccaggca tcagggacgt ggccgtcgtg ggtatccctg acatcgaggc
     2101 cggcgagctg ccagccggct tcgtggtgaa gcagcccggc gcccaactca ccgctaagga
     2161 ggtgtacgac ttcctggccc agagggtgtc tcactccaag tacctgaggg gcggcgtaag
     2221 gttcgtggac tctatcccca ggaacgtgac aggcaagatt agtcgaaaag agctgaggga
     2281 ggccctgatg gagaaggctt ctaagctgct cgaggactac aaagaccatg acggtgatta
     2341 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     2401 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2461 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2521 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2581 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2641 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2701 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2761 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     2821 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2881 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     2941 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     3001 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3061 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3121 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3181 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3241 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     3301 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3361 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3421 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     3481 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3541 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3601 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3661 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3721 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3781 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3901 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3961 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4021 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4081 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4141 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4201 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4261 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4321 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4381 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4441 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4501 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4561 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4621 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4681 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4741 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4801 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4861 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4921 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     4981 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5041 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5101 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5161 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5221 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5401 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5521 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5881 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6121 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6601 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7441 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     7501 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7561 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     7621 tgcttagg
//
