LOCUS       62056_19830        9731 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 9731)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5228..5293
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    5331..5555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5601..6029
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6043..6372
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6439..7230
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    7407..7540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7577..7593)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7601..7617)
                     /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(7625..7655)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7670..7691)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7979..8564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8738..9595)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9596..9700)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgagtatgc taaagccaag
      961 tgggcttaag gcccccacca agatcctgaa gcctggaagc acagctctga agacacctac
     1021 ggctgttgta gctccagtag aaaaaaccat atccagtgaa aaagcatcaa gcactccatc
     1081 atctgagact caggaggaat ttgtggatga ctttcgagtt ggggagcgag tttgggtgaa
     1141 tggaaataag cctggattta tccagtttct tggagaaacc cagtttgcac caggccagtg
     1201 ggctggaatt gttttagatg aacccatagg caagaacgat ggttcggtgg caggagttcg
     1261 gtatttccag tgtgaacctt taaagggcat atttacccga ccttcaaagt taacaaggaa
     1321 ggtgcaagca gaagatgaag ctaatggcct gcagacaacg cccgcctccc gagctacttc
     1381 accgctgtgc acttctacgg ccagcatggt gtcttcctcc ccctccaccc cttcaaacat
     1441 ccctcagaaa ccatcacagc cagcagcaaa ggaaccttca gctacgcctc cgatcagcaa
     1501 ccttacaaaa actgccagtg aatctatctc caacctttca gaggctggct caatcaagaa
     1561 aggagaaaga gagctcaaaa tcggagacag agtattggtt ggtggcacta aggctggtgt
     1621 agtccggttt cttggggaga ccgactttgc caagggggag tggtgtggcg tggagttaga
     1681 tgagccactt gggaagaatg atggcgctgt tgctggaaca aggtattttc agtgtcaacc
     1741 caaatatggc ttgttcgctc ctgtccacaa agttaccaag attggcttcc cttccactac
     1801 accagccaaa gccaaggcca acgcagtgag gcgagtgatg gcgaccacgt ccgccagcct
     1861 gaagcgcagc ccttctgcct cttccctcag ctccatgagc tcagtggcct cctctgtgag
     1921 cagcaggccc agtcggacag gactattgac tgaaacctcc tcccgttacg ccaggaagat
     1981 ctccggtacc actgccctcc aggaggccct gaaggagaag cagcagcaca ttgagcagct
     2041 gctggcggaa cgggatctgg agagggcgga ggtggccaag gccacgagcc acgtggggga
     2101 gatagagcag gagctagctc tggcccggga cggacatgac cagcatgtcc tggaattgga
     2161 agccaaaatg gaccagctgc gaacaatggt ggaagctgct gacagggaga aggtggagct
     2221 tctcaaccag cttgaagagg agaaaaggaa ggttgaggac cttcagttcc gggttgaaga
     2281 agaatcaatt accaaaggtg atcttgagac gcagaccaaa ctggagcatg cccgcattaa
     2341 ggagcttgaa cagagcctgc tctttgaaaa gaccaaagct gacaaactcc agagggagtt
     2401 agaagacact agggtggcta cagtttcaga aaagtcacgt ataatggaac tggagaaaga
     2461 cctagcattg agagtacagg aagtagctga gctccgaaga aggctagagt ccaataagcc
     2521 tgctggggat gtggacatgt cactttccct tttgcaagag ataagctctt tgcaagaaaa
     2581 gttagaagtc acccgtactg accaccagag agaaataact tctctgaagg agcattttgg
     2641 agcccgggaa gaaactcatc agaaggagat aaaggctctg tataccgcca cggaaaagct
     2701 ttccaaagag aacgagtcat tgaaaagcaa gctggagcat gccaacaaag agaactcaga
     2761 tgtgatagct ctatggaagt ccaaactgga gactgccatc gcatcccacc agcaggcgat
     2821 ggaagaactg aaggtatctt tcagcaaagg gcttggaaca gagacggcag aatttgctga
     2881 actaaaaaca caaatagaga aaatgagact agattaccaa cacgaaatag aaaatttgca
     2941 gaatcaacaa gactctgaac gggctgccca tgctaaagag atggaagcct tgagggctaa
     3001 actgatgaaa gttattaaag aaaaggaaaa cagtctggaa gccatcaggt cgaaactgga
     3061 caaagcagaa gaccagcatc tcgtagaaat ggaagacacg ttaaacaaat tacaggaagc
     3121 tgaaataaag gtaaaggagc tagaggtact gcaagccaaa tgcaatgaac aaaccaaggt
     3181 tattgataat tttacatcac agctcaaggc tactgaagaa aagctcttgg atcttgatgc
     3241 acttcggaaa gccagttccg aaggtaaatc ggaaatgaag aaacttagac agcagcttga
     3301 ggcagctgag aaacagatta aacatttaga gattgaaaag aatgctgaaa gtagcaaggc
     3361 tagtagcatt accagagagc tccaggggag agagctaaag cttactaacc ttcaggaaaa
     3421 tttgagtgaa gtcagtcaag tgaaagagac tttggaaaaa gaacttcaga ttttgaaaga
     3481 aaagtttgct gaagcttcag aggaggcagt ctctgttcag agaagtatgc aagaaactgt
     3541 aaataagtta caccaaaagg aggaacagtt taacatgctg tcttctgact tggagaagct
     3601 gagagaaaac ttagcagata tggaggcaaa atttagagag aaagatgaga gagaagagca
     3661 gctgataaag gcaaaggaaa aactggaaaa tgacattgca gaaataatga agatgtcagg
     3721 agataactct tctcagctga caaaaatgaa cgatgaatta cgtctgaaag aaagagatgt
     3781 agaagaatta cagctaaaac ttacaaaggc taatgaaaat gcaagttttc tgcaaaaaag
     3841 tattgaggac atgactgtca aagctgaaca gagccagcaa gaagcagcta aaaagcatga
     3901 ggaagaaaag aaagaattgg agaggaaatt gtcggacctg gaaaagaaaa tggaaacaag
     3961 ccacaaccag tgtcaggagc tgaaagccag gtatgagaga gccacttctg agacaaaaac
     4021 caagcatgaa gaaatcctac agaacctcca gaagacgctg ctggacacag aggacaagct
     4081 gaagggcgca cgggaggaga acagtggctt gctgcaggag ctggaggagc tgagaaagca
     4141 agccgacaaa gccaaagctg ctcaaacagc ggaagatgcc atgcagataa tggaacagat
     4201 gaccaaagag aagactgaga ctctggcctc cttggaggac accaagcaaa caaatgcaaa
     4261 actacagaat gaattggaca cacttaaaga aaacaacttg aaaaatgtgg aagagctgaa
     4321 caaatcaaaa gaactcctga ctgtagagaa tcaaaaaatg gaagaattta ggaaagaaat
     4381 agaaacccta aagcaggcag cagctcagaa gtcccagcag ctttcagcgt tgcaagaaga
     4441 gaacgttaaa cttgctgagg agctggggag aagcagggac gaagtcacaa gtcatcaaaa
     4501 gctggaagaa gaaagatctg tgctcaataa tcagttgtta gaaatgaaaa aaagagaatc
     4561 caagttcata aaagacgcag atgaagagaa agcttccttg cagaaatcca tcagtataac
     4621 tagtgcctta ctcacagaaa aggatgccga gctggagaaa ctgagaaatg aggtcacagt
     4681 gctcagggga gaaaacgcct ctgccaagtc cttgcattca gttgttcaga ctctagagtc
     4741 tgataaggtg aagctcgagc tcaaggtaaa gaacttggag cttcaactca aagaaaacaa
     4801 gaggcagctc agcagctcct caggtaatac agacactcag gcagacgagg atgaaagagc
     4861 ccaggagagt cagattgatt tcctaaattc agtaatagtg gaccttcaaa ggaagaatca
     4921 agacctcaag atgaaggtgg agatgatgtc agaagcagcc ctgaatggga acggggatga
     4981 cctaaacaat tatgacagtg atgatcagga gaaacagtcc aagaagaaac ctcgcctctt
     5041 ctgtgacatt tgtgactgct ttgatctcca cgacacagag gattgtccta cccaggcaca
     5101 gatgtcagag gaccctcccc attccacaca ccatggcagt cggggtgagg aacgcccata
     5161 ctgtgaaatc tgtgagatgt ttggacactg ggccaccaac tgcaatgacg acgaaacctt
     5221 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     5281 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     5341 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5401 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5461 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5521 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5581 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5941 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6001 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     6061 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6121 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6181 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6241 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6301 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6361 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     6421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6601 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6961 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     7021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     7081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7261 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     7321 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     7381 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     7441 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7501 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     7561 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7621 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7681 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7741 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7801 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     8041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     8101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8401 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     8521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8821 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8881 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8941 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     9001 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     9061 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     9121 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9181 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9241 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9301 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9361 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9421 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9481 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9541 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9601 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9661 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9721 cacctgacgt c
//
