LOCUS       62056_9355        6049 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 6049)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6049
                     /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             739..804
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    825..1413
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    1479..1527
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1729..2157
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2171..2500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2567..3358
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3537..3670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3707..3723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3731..3747)
                     /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(3755..3785)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3800..3821)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4109..4697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4871..5728)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5729..5833)
                     /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 gcgttttcta gaaaacttaa
      661 gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattctgcag atatccagca
      721 cagtggcggc cgctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc
      781 gactacaagg atgacgatga caagtagtga gggcccatct cgacaatcaa cctctggatt
      841 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
      901 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
      961 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     1021 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     1081 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     1141 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     1201 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     1261 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     1321 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     1381 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac
     1441 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     1501 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     1561 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     1621 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     1681 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     1741 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1801 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1861 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc
     1921 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1981 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2041 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg
     2101 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2161 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2221 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2281 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2341 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2401 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2461 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2521 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2581 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2641 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2701 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2761 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2821 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2881 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2941 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3001 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3061 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3121 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3181 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3241 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3301 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3361 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3421 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3481 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3541 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3601 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3661 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3721 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3781 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3841 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3901 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     4321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4861 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4921 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4981 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5041 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5101 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5161 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5221 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5281 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5341 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5401 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5461 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5521 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5581 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5641 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5701 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5761 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5821 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     5881 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
     5941 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6001 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
