LOCUS       62056_7515        6758 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 6758)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6758
                     /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             1447..1464
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     misc_feature    1534..2122
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    2188..2236
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2438..2866
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2880..3209
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3276..4067
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4246..4379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4416..4432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4440..4456)
                     /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(4464..4494)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4509..4530)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4818..5406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5580..6437)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6438..6542)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatgcgg
      781 gtgcagccca ccgaatccat cgtgcggttc cccaatatca ccaatctgtg ccccttcgat
      841 gaggtgttca atgccaccag attcgccagc gtgtacgcct ggaaccggaa gagaatcagc
      901 aactgcgtgg ccgactactc cgtgctgtac aatctggccc cattcttcac cttcaagtgc
      961 tacggcgtgt cccctaccaa gctgaacgac ctgtgcttca ccaatgtgta cgccgacagc
     1021 ttcgtgatcc ggggagatga agtgcggcag attgcccctg gacagaccgg caatatcgcc
     1081 gactacaact acaagctgcc cgacgacttc accggctgtg tgatcgcctg gaatagcaac
     1141 aagctggaca gcaaggtgtc cggcaactac aattacctgt accggctgtt ccggaagtcc
     1201 aatctgaagc ccttcgagcg ggacatcagc accgaaatct atcaggccgg caacaagccc
     1261 tgcaatggcg tggcaggctt caactgctac ttcccactgc ggagctacag cttcagaccc
     1321 acatacggcg ttggccacca gccttacaga gtggtggtgc tgtccttcga gctgctgcat
     1381 gctcctgcca cagtgtgcgg ccctaagaaa agcaccaacc tcgtgaagaa caaatgcgtg
     1441 aacttccacc atcaccatca ccattagctc gagtctagag ggccccttaa gggttcgatc
     1501 cctaccggtt agtaatgagt ttgatatctc gacaatcaac ctctggatta caaaatttgt
     1561 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1621 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1681 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     1741 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     1801 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     1861 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     1921 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     1981 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2041 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2101 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     2161 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     2221 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     2281 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     2341 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     2401 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2461 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2521 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2581 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2641 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2701 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2761 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     2821 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2881 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2941 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3001 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3061 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3121 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3181 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3241 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3301 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3361 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3421 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3481 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3541 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3601 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3661 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3721 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3781 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3841 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3901 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3961 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4021 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4081 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4141 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4201 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4261 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4321 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4381 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4441 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4501 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4561 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4621 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4681 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4741 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4981 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     5041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5221 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5341 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5401 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5461 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5521 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5581 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5641 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5701 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5761 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5821 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5881 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5941 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6001 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6061 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6121 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6181 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6241 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6301 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6361 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6421 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6481 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6541 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     6601 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     6661 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     6721 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
