LOCUS       62056_1990        7884 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 7884)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             931..996
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2643..3265
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        3307..3604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3658..4449
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5104..5238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5244..5262)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5276..5292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5300..5316)
                     /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(5324..5354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5369..5390)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5678..6266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6440..7297)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7298..7402)
                     /label=AmpR promoter
     rep_origin      7429..7884
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcaatgaac
     1021 gaggaggcca tctgcagcgc cctgcccacc atcccgtacc gaaagctcgc tgacctgtgc
     1081 tacttgagcc gtggcgcgtc cggcaccgtg tcgtccgccc gccacgcaga ctggcgcatc
     1141 caggtggccg taaagcactt gcacatccac accccactgc ttgacagtga aagaaatgat
     1201 gtcttaagag aagctgaaat tttacacaaa gctagattta gttacattct tccaattttg
     1261 ggaatttgca atgagcctga atttttggga atagttaccg aatatatgcc aaatggatcg
     1321 ttaaatgaac tcctacatag gaaaaatgaa tatcctgatg ttccttggcc tttgagattt
     1381 cgtattttgc atgaaattgc actaggtgta aattacctgc acaatatgaa tcctccttta
     1441 cttcaccatg atctgaagac tcaaaatatc ttattggata atgaatttca tgttaagatt
     1501 gcagattttg gtttatcaaa atggcgtatg atgtccctct cacaatcacg aagtattaaa
     1561 tctgctccag aaggagggac aattatctat atgccacctg aaaactatga acctggacaa
     1621 aaatcaaggg ctactgtcaa gcatgatata tatagttatg cagttatcac atgggaagtc
     1681 ttatccagaa aacagccttt tgaagaagtc accaatcctt tgcagattat gtatagcgtg
     1741 tcacaaggac atcgacctga cactagcgaa gaaagtttgc cgtttgatat acctcatcga
     1801 gcactgatga tctctctaat agaaagtgga tgggcacaaa atccagatga aagaccatct
     1861 ttcttaaaat gtttaataga acttgaacct gttctgagaa tatttgaaga tataactttc
     1921 cttgaagctg ttattcaact aaagaaaaca aagttgcaaa gtgcttcgag tactgttcac
     1981 ttatgtgaca agaagaaaac agagttatat ccgaacacat ctgtaaatca tggttcacaa
     2041 gaggaatcat gtggatcttc gcagcttcat aaacctagta attctccagg aacctcaaag
     2101 tccctgtcag ctactcaaga cagtgacttt ttatctagaa aaactcaaga cttttccacg
     2161 ccacgtcagt gtccagtaaa tcgcagttgg gacagtaaca tttctgtggc tcaaagggca
     2221 acattctgtg atcacaagac caccccatgc tctttagcaa taataaatcc actctcagcc
     2281 gaaggaaatt cagagcgatt tcagcctggt atagcccaac agtggatcca gagtaaaagg
     2341 gaagacattg tgaaccagat gacggaagcc tgtcttaacc aatctctgga tgccctcctg
     2401 tccagggatt tgatcatgaa ggaggactat gaactcatta gtaccaaacc tacaagaacc
     2461 tcaaaagtca gacaattact ggataccact gacatccaag gagaagaatt tgccagagtt
     2521 atagtacaaa agttgaaaga taacaagcaa atgggactcc agccttaccc agaaatactt
     2581 gtggtttctc gaccaccatc tttaaattta ttttagggta ccagtcgact ctagaggatc
     2641 ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     2701 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     2761 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     2821 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     2881 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     2941 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     3001 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     3061 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     3121 tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc
     3181 atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt
     3241 ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc
     3301 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3361 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3421 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3541 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3601 ttggaggaat tgatcagctt gggatctgat caagagacag gatgaggatc gtttcgcatg
     3661 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3721 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3781 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3841 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3901 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3961 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4021 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4081 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4141 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4201 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4261 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4321 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4381 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4441 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4501 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4561 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4621 accccgggct cgatcccctc gcgagttggt tcagctgctg cctgaggctg gacgacctcg
     4681 cggagttcta ccggcagtgc aaatccgtcg gcatccagga aaccagcagc ggctatccgc
     4741 gcatccatgc ccccgaactg caggagtggg gaggcacgat ggccgctttg gtcgacccgg
     4801 acgggacgct cctgcgcctg atacagaacg aattgcttgc aggcatctca tgagtgtgtc
     4861 ttcccgtttt ccgcctgagg tcactgcgtg gatggagcgc tggcgcctgc tgcgcgacgg
     4921 cgagctgctc accacccact cgccaagctg gaaccgtaaa aaggccgcgt tgctggcgtt
     4981 tttccatagg ctccgccgat cataatcagc cataccacat ttgtagaggt tttacttgct
     5041 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     5101 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5221 tcttatcatg tctggatcaa ttccctatag tgagtcgtat taaattcgta atcatgtcat
     5281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5341 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5461 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5521 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6181 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6241 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6301 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6361 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6421 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6481 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6541 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6601 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6661 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6721 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6781 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6841 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6901 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6961 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7021 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7081 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7141 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7201 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7261 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7321 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7381 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     7441 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7501 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7561 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7621 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7681 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7741 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7801 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     7861 acaaaatatt aacgcttaca attt
//
