LOCUS       62056_23440        8182 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 8182)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..47
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        57..75
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     158..174
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3383..3686
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3687..3890
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     polyA_signal    4142..4263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4387..4890
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4925..5947
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(6017..6035)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6056..6072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6080..6096)
                     /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(6104..6134)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6149..6170)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6458..7046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7220..8077)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8078..8182)
                     /label=AmpR promoter
ORIGIN
        1 gcggccgcca gtggaacgaa aactcacgtt gtaaaacgac ggccagtgag cgcgcgtaat
       61 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaagtttat ctagggataa
      121 cagggtaatg ataaacattt cggataacaa tttcacacag gaaacagcta tgaccatgat
      181 tacgccaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt
      241 cgccctttac cactaagcca tctctccagc cctgagtcat ggttttagtg tgagaggcat
      301 cattgaattt tctgagcacg gccatcaggg tagctggcac aggtcttcag atacaaggag
      361 atagttataa gaaggcagcc atggctgtgg tgcactagaa atggagaaac agcttcatca
      421 ggtgacagac cagtctgact ctgtcccatg attagaagcc atcttgttac aaggtcaaaa
      481 taagttcatt cctgttttct gtaacacttg ggtttgatcc tgtcgtcaac ccattttctg
      541 gaatttgaca tgttccatac tccattatac cctgacttcc accctgataa gatgttctgc
      601 caagttcctg tgtagccaac attcccctgg aaatctctct tcccttggaa accacctagt
      661 cttagaaatt ttgagttata taaattccac ttctatgttt gatgctattc tttaaaactc
      721 cactttaggg agatagccct gtctgataga aaataaaact tgcttaattt gtctaaaaga
      781 ttttaagtaa tagtttttac ttttgttccg tgggattagt acagggtgaa acagactccc
      841 gtgtttccag tgtgaagtga gccacacact gcagtacaag ttatatcagc aggttctgcc
      901 tctgcgcaat gaacttttgc ttgtgtggac atcagggtct gtgtgaaggg aaggtcctat
      961 ggcctagttt tatactattc aacagtctgt ccccgaagcc ctggtgcttt attattttga
     1021 caagcccctg ctgctggtat tccaccctgc tgcgagtcaa aaaagttcct gtctcggaaa
     1081 aacaaaacaa aacaaaacaa ccaaaaaata aatttttttt tcccacaggt tctagtggag
     1141 gtgctcacta ccagaaatcc tacaaataag cccatctcat ggatcagggt ttacctttgt
     1201 aataatatta aatctgtgtg catgtgcgca cgcatgtgtt ttatgcttgc atatatgtat
     1261 acgcagccat ggttttctac tgtcccactc actctgtaac ttactgagcc atccagctgg
     1321 tcctctaaat acatttcaat gaaagttttc attagcgtga acgtgaaggt ggtaaaatct
     1381 gttagtgtgt gcttatgcct gtggtttgca cctctagtct gaaggttgct cttttcaaat
     1441 tttttattta tttacgtttt tacttctgag tcagaaactc ataaaggcca tggcctcgaa
     1501 ttcgctatgt agtcaacgat gaccttaaac ttgtgaccct ctacttcgtt agtgctggaa
     1561 ccccaagctt gctgagtaca gagcactttc agaccggaac tagatgtcta cttcctgttc
     1621 cgcctacatt acaggttgct aggttacacc ccccctacgc cgttttagac gcaaaacttc
     1681 atttcccatg caaaacttca tttcccatga acacttgcaa gggtcgccgc gctgcgcggc
     1741 gtcattgctc ccgccctata tacctacttc cgcccgcgag ccacttcctt tcctttcagc
     1801 ggcgcgcggc tgcaagatgg cggtgcagat ttccaagaag aggaaggtaa gcgtctgggc
     1861 ccggttcggg agtccgccgc gggttctaca agtgccaggg aggcctgtgg ctccgtaatc
     1921 agtcctgtgg agcgtctggg gccgcctgcc gtctcttcga gcctcggatg gccgtagatt
     1981 gtgtattggg ccggagccgg gcgagtgctg tgtgcctggg caagggaggg acaaactcct
     2041 cgagttctgg accgactcga acaccgggcg cctccagttc cggactagac acctttgagc
     2101 gtttcttggt ctccataata gtaatcctgt ggcacagtta gagggcgtgt gccatcagat
     2161 ctagtccagt ctctttagta agtgaagttt agcagtccct tctcttagtc gcgtgatcct
     2221 gcaagtggcc atagttgaaa gcctacttac tgactgctgc cgtgttcact cgggacccgg
     2281 agctgcagcg tccctgtggt tatcatttca tgggggaaaa gtgtgcaggt tgccaggttt
     2341 agaaatagat ggtctgtcgt ttgtgcttat gcacacagat gataaacctg ttttgagtca
     2401 ggattcctct cctatccgag gtacaactta cagtcccagc tgtacatgtg ctacttggag
     2461 acagattttt ctttgtctct tgggtgtaga ttatgccgta gagcccttcg atgaagaggt
     2521 gatgacgagt ctgagtagga agtgttgtct ttgtccaaga tgcctcacta tgctgcgttc
     2581 tgtggcacag ctgaaagcac tgtggtcaaa agaaacttcc taaagatgac caagaggcat
     2641 ttgtctgaga agggttgctg cttttctgta gggccattgg gcttgctctg actaaccctg
     2701 tcttcacctc agaggtaact tgtttccttt ggttcagttt gtagctgatg gcatcttcaa
     2761 agctgagctg aatgaatttc tcactcggga gctggctgaa gatggctact ctggagttga
     2821 agtccgagtt acaccaacca ggacagaaat cattatttta gccaccaggt agaaatacca
     2881 ttgattgtca cctgtaaata ttgtgtgtac tgagatgctg tgtaaacttg ggccaaccaa
     2941 gcagtaaatc tggcctcagt gggtgtaact gctttgttag aactgcattt gggaagaact
     3001 taccttccat ttaacctgtg tgctggcgtt gtggtgggcg gcaggtggga tcttgagtaa
     3061 atggttgcgc ttcccctcta caggacacag aatgttcttg gggagaaggg tcgtcggatc
     3121 agagagttga ccgcagttgt ccagaagcgc tttggcttcc ctgaaggcag cgtagaggtg
     3181 agttcctctg ctttatctcc cgggggtttt agactgagtt gggatgtggc ttctgctata
     3241 gaattgtact tctgaaaacc tgacatggcc agtgacagtc acaggtactt gatgctaagg
     3301 gcgaattctg cagatgggcc gtacgaattt aatagtaatc aattacgggg tcattagttc
     3361 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3421 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3481 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3541 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     3601 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     3661 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     3721 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     3781 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     3841 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     3901 accgtcagat ccgctagaat tcgagctccc tgcaggttag ttaagttaac ggccggccgc
     3961 tagcgcgatc gcgagctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag
     4021 ctggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4081 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4141 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4261 ttaaagctat ccatcacact ggcggccgaa atctagaagc ttatcgatat ggcgcgccaa
     4321 ttgttaggcc gcattcgcca atcaagtaag ctaagatctg gcccaatggg ccgtacgaat
     4381 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     4441 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     4501 gccggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     4561 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     4621 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     4681 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     4741 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     4801 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     4861 ttctcctctt cctcatctcc gggcctttcg acggtaccgc gggcccgggg gatccactag
     4921 tactatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     4981 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     5041 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     5101 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     5161 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     5221 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctc cagccggtcg cggaggccat
     5281 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     5341 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     5401 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     5461 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc atgcggattt
     5521 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     5581 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ctcttctgga ggccgtggtt
     5641 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     5701 gccgcgcctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     5761 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     5821 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     5881 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     5941 aaaggaatag agtagatgcc gaccgggaac cagttaacgt cgacctcgag ggggggcccg
     6001 gtaccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat
     6061 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6121 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6181 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6241 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6301 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6361 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6421 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6481 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6541 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6601 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6661 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6721 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6781 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6841 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6901 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6961 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7021 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7081 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7141 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7201 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7261 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7321 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7381 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7441 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7501 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7561 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7621 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7681 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7741 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7801 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7861 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7921 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7981 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8041 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8101 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8161 ataaacaaat aggggttccg cg
//
