LOCUS       62056_17055        6997 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 6997)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6997
                     /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             664..729
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1773..2361
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    2427..2475
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2677..3105
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3119..3448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3515..4306
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4485..4618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4655..4671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4679..4695)
                     /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(4703..4733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4748..4769)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5057..5645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5819..6676)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6677..6781)
                     /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 ttaagccacc
      661 atggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
      721 gatgacaagc ttggtaccga gctcggatcc atgaaggaga ataaagaaaa ttcaagccct
      781 tcagtaactt cagcaaacct ggaccacaca aagccatgtt ggtactggga taagaaagac
      841 ttggctcata caccctcaca acttgaagga cttgatccag ccaccgaggc ccggtaccgc
      901 cgagagggcg ctcggttcat ctttgatgtg ggcacacgtt tggggctaca ctatgatacc
      961 ctggcaactg gaataattta ttttcatcgc ttctatatgt ttcattcctt caagcaattc
     1021 ccaagatatg tgacaggagc ctgttgcctc tttctggctg ggaaagtaga agaaacacca
     1081 aaaaaatgta aagatatcat caaaacagct cgtagtttat taaatgatgt acaatttggc
     1141 cagtttggag atgacccaaa ggaggaagta atggttctgg agagaatctt actgcagacc
     1201 atcaagtttg atttacaggt agaacatcca taccagttcc tactaaaata tgcaaagcaa
     1261 ctcaaaggtg ataaaaacaa aattcaaaag ttggttcaaa tggcatggac atttgtaaat
     1321 gacagtctct gcaccacctt gtcactgcag tgggaaccag agatcatagc agtagcagtg
     1381 atgtatctcg caggacgttt gtgcaaattt gaaatacaag aatggacctc caaacccatg
     1441 tataggagat ggtgggagca gtttgttcaa gatgtcccgg tcgacgtttt ggaagacatc
     1501 tgccaccaaa tcctggatct ttactcacaa ggaaaacaac agatgcctca tcacaccccc
     1561 catcagctgc aacagccccc atctcttcag cctacaccac aagtgccgca agtacagcag
     1621 tcacagccgt ctcaaagctc cgaaccatcc cagccccagc agaaggaccc ccagcaacca
     1681 gcccagcagc agcagggcat gcctccagtt ggggggctgg ggcgggcagc ctggatgaga
     1741 taactcgagt ctagagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     1801 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1861 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1921 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1981 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2041 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2101 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2161 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2221 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2281 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2341 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     2401 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     2461 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     2521 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     2581 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     2641 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2701 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2761 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2821 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2881 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2941 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3001 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     3061 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3121 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3181 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3241 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3301 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3361 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     3421 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     3481 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3541 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3601 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3661 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     3721 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3781 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3841 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3901 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3961 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4021 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4081 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4141 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4201 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4261 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4321 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4381 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4441 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4501 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4561 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4621 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4681 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4741 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4801 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4861 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4921 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4981 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5041 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5101 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5161 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5221 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     5281 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5341 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5401 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5461 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5521 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5581 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5641 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5701 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5761 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5821 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5881 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5941 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6001 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6061 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6121 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6181 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6241 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6301 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6361 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6421 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6481 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6541 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6601 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6661 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6781 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     6841 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     6901 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     6961 aggcttgacc gacaattgca tgaagaatct gcttagg
//
