LOCUS       62056_13500        8721 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 8721)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..34
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        115..143
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    151..167
                     /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)."
     CDS             200..613
                     /codon_start=1
                     /label=Gam
                     /note="inhibitor of the host RecBCD nuclease in the lambda 
                     Red system"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             622..1404
                     /codon_start=1
                     /label=Beta
                     /note="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             1404..2081
                     /codon_start=1
                     /label=Exo
                     /note="5' to 3' double-stranded DNA exonuclease in the 
                     lambda
                     Red system"
                     /translation="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     primer_bind     complement(2118..2134)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2152..2229
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2230..3309
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    3325..3346
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3361..3391
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     primer_bind     3423..3439
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3455..3471)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3608..3626)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3647..3663)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3671..3687)
                     /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(3695..3725)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3740..3761)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5236..6183)
                     /codon_start=1
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(6231..6453)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(7084..7941)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7942..8046)
                     /label=AmpR promoter
     rep_origin      complement(8072..8527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8669..8685
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8695..8713
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag
       61 cccgggggat ctccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca
      121 attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      181 acagtcgacg cttataaaaa tggatattaa tactgaaact gagatcaagc aaaagcattc
      241 actaaccccc tttcctgttt tcctaatcag cccggcattt cgcgggcgat attttcacag
      301 ctatttcagg agttcagcca tgaacgctta ttacattcag gatcgtcttg aggctcagag
      361 ctgggcgcgt cactaccagc agctcgcccg tgaagagaaa gaggcagaac tggcagacga
      421 catggaaaaa ggcctgcccc agcacctgtt tgaatcgcta tgcatcgatc atttgcaacg
      481 ccacggggcc agcaaaaaat ccattacccg tgcgtttgat gacgatgttg agtttcagga
      541 gcgcatggca gaacacatcc ggtacatggt tgaaaccatt gctcaccacc aggttgatat
      601 tgattcagag gtataaaacg aatgagtact gcactcgcaa cgctggctgg gaagctggct
      661 gaacgtgtcg gcatggattc tgtcgaccca caggaactga tcaccactct tcgccagacg
      721 gcatttaaag gtgatgccag cgatgcgcag ttcatcgcat tactgatcgt tgccaaccag
      781 tacggcctta atccgtggac gaaagaaatt tacgcctttc ctgataagca gaatggcatc
      841 gttccggtgg tgggcgttga tggctggtcc cgcatcatca atgaaaacca gcagtttgat
      901 ggcatggact ttgagcagga caatgaatcc tgtacatgcc ggatttaccg caaggaccgt
      961 aatcatccga tctgcgttac cgaatggatg gatgaatgcc gccgcgaacc attcaaaact
     1021 cgcgaaggca gagaaatcac ggggccgtgg cagtcgcatc ccaaacggat gttacgtcat
     1081 aaagccatga ttcagtgtgc ccgtctggcc ttcggatttg ctggtatcta tgacaaggat
     1141 gaagccgagc gcattgtcga aaatactgca tacactgcag aacgtcagcc ggaacgcgac
     1201 atcactccgg ttaacgatga aaccatgcag gagattaaca ctctgctgat cgccctggat
     1261 aaaacatggg atgacgactt attgccgctc tgttcccaga tatttcgccg cgacattcgt
     1321 gcatcgtcag aactgacaca ggccgaagca gtaaaagctc ttggattcct gaaacagaaa
     1381 gccgcagagc agaaggtggc agcatgacac cggacattat cctgcagcgt accgggatcg
     1441 atgtgagagc tgtcgaacag ggggaagatg cgtggcacaa attacggctc ggcgtcatca
     1501 ccgcttcaga agttcacaac gtgatagcaa aaccccgctc cggaaagaag tggcctgaca
     1561 tgaaaatgtc ctacttccac accctgcttg ctgaggtttg caccggtgtg gctccggaag
     1621 ttaacgctaa agcactggcc tggggaaaac agtacgagaa cgacgccaga accctgtttg
     1681 aattcacttc cggcgtgaat gttactgaat ccccgatcat ctatcgcgac gaaagtatgc
     1741 gtaccgcctg ctctcccgat ggtttatgca gtgacggcaa cggccttgaa ctgaaatgcc
     1801 cgtttacctc ccgggatttc atgaagttcc ggctcggtgg tttcgaggcc ataaagtcag
     1861 cttacatggc ccaggtgcag tacagcatgt gggtgacgcg aaaaaatgcc tggtactttg
     1921 ccaactatga cccgcgtatg aagcgtgaag gcctgcatta tgtcgtgatt gagcgggatg
     1981 aaaagtacat ggcgagtttt gacgagatcg tgccggagtt catcgaaaaa atggacgagg
     2041 cactggctga aattggtttt gtatttgggg agcaatggcg atgacgcatc ctcacgataa
     2101 tatccgggta ggcgcaagat ccactagttc tagagcggcc gctaattcgg ggacaccatc
     2161 gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     2221 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     2281 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     2341 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     2401 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     2461 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     2521 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     2581 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     2641 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     2701 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     2761 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     2821 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     2881 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     2941 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     3001 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     3061 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     3121 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     3181 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     3241 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3301 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc
     3361 tttacacttt atgcttccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     3421 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
     3481 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3541 gctggcgtaa tagcgaagag gcccgcaccg agcggccgcc accgcggtgg agctccagct
     3601 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     3661 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3721 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3781 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3841 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3901 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3961 cagaatcagg ggataacgca ggaaagaaca tgtgagatcc ccacagcaag atgcaacttt
     4021 cgccagatac gacggcgctc tttgccgtgc tttctgactt tccattcgcc ttcaccaaag
     4081 accttcagcc cggtggaatc aatcaccagg tgtgcgattt caccccgggt ggacgttttg
     4141 aaactgacat taaccgactt tgcccgctta ctgacactgg tgtaatccgg gcagcgcaac
     4201 ggaacgttca tcagggcaaa aatggaatca ataaaaccct gcgcagcccg cagggtcagc
     4261 cggaatacgc gtttaatcac cagaacggtg gtgatggcga gatcagaata gcgctggggc
     4321 cttcctcgtg atgaaggtgt tgccgactca taccaggcct gaatcgcctc atcatccagc
     4381 cagaaagtga gggagccacg gttgatgaga gctttgttgt aggtggacca gttggtgatt
     4441 ctgaactttt gctttgccac ggaatggtct gtgttgtcgg gaggatgcgt gatctgatcc
     4501 ttcaactcag caaaagttcg atttattcaa caaagccact ttcagtatgg atttgggtgt
     4561 tacggggggg ggttggtgct taaaatcgcg ctgaggattt ctgatggtgt taagcgggcg
     4621 gttttgagat gtaaactcgc ccgtttaaca taatggatct tgcgcgcacc gcccgaacac
     4681 cactcgccac aaaaaaccgc cggaacgtcc aaaagtacgg gttttgctgc ccgcaaacgg
     4741 gctgttctgg tgttgctagt ttgttatcag aatcgcagat ccggcttcag ccggtttgcc
     4801 ggctgaaagc gctatttctt ccagaattgc catgattttt tccccacggg aggcgtcact
     4861 ggctcccgtg ttgtcggcag ctttgattcg ataagcagca tcgcctgttt caggctgtct
     4921 atgtgtgact gttgagctgt aacaagttgt ctcaggtgtt caatttcatg ttctagttgc
     4981 tttgttttac tggtttcacc tgttctatta ggtgttacat gctgttcatc tgttacattg
     5041 tcgatctgtt catggtgaac agctttaaat gcaccaaaaa ctcgtaaaag ctctgatgta
     5101 tctatctttt ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatgtaa
     5161 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     5221 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     5281 tttttgcgtg agccatgaga acgaaccatt gagatcatac ttactttgca tgtcactcaa
     5341 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     5401 tcttagtccg ttatgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     5461 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     5521 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     5581 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     5641 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     5701 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     5761 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     5821 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     5881 ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac
     5941 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     6001 gatgttcatc atctgaacgt attggttata agtgaacgat accgtccgtt ctttccttgt
     6061 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     6121 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     6181 catacatctc aattggtcta ggtgatttta atcactatac caattgagat gggctagtca
     6241 atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt
     6301 tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt
     6361 tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat
     6421 aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta
     6481 caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg
     6541 cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca
     6601 tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg
     6661 cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac
     6721 ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat
     6781 gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac
     6841 cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg
     6901 gtatcatcaa caggcttacc cgtcttactg tcggggatct gacgctcagt ggaacgaaaa
     6961 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7081 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7141 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7201 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7261 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7321 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7381 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7441 cagctccggt tcccagcgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7501 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7561 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7621 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7681 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7741 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7801 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7861 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7921 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7981 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8041 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     8101 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     8161 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     8221 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     8281 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     8341 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     8401 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     8461 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     8521 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8581 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8641 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     8701 gactcactat agggcgaatt g
//
