LOCUS       62056_22345        7889 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 7889)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(891..1242)
                     /direction=LEFT
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             1256..2086
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     primer_bind     2236..2252
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2266..3141)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3168..3452
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             3496..3909
                     /codon_start=1
                     /label=Gam
                     /note="inhibitor of the host RecBCD nuclease in the lambda 
                     Red system"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             3918..4700
                     /codon_start=1
                     /label=Beta
                     /note="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             4700..5377
                     /codon_start=1
                     /label=Exo
                     /note="5' to 3' double-stranded DNA exonuclease in the 
                     lambda
                     Red system"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      5381..5625
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     primer_bind     complement(5668..5684)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5692..5708)
                     /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(5716..5746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5761..5782)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6070..6658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6832..7689)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7690..7794)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcggc
      241 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
      301 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
      361 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat
      421 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
      481 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
      541 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
      601 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
      661 cccttaacgt gagttttcgt tccactgagc gtcagacccc aattacacgc cactggctgt
      721 gcttgctggg gtgacggtgg caacggtggc ggccttgctg ggctatcgcg ttggaaagaa
      781 acgagggaaa ggggactgat aaaccggtct tagcccctcc ccttggtgtc caaccgctct
      841 gtaggcctct caggcgccgc tggtgccgct ggttggacgc caagggtgaa tccgcctcga
      901 taccctgatt actcgcttcc tgcgccctct caggcggcga taggggactg gtaaaacggg
      961 gattgcccag acgcctcccc cgccccttca ggggcacaaa tgcggcccca acggggccac
     1021 gtagtggtgc gttttttgcg tttccaccct tttcttcctt ttccctttta aaccttttag
     1081 gacgtctaca ggccacgtaa tccgtggcct gtagagttta aaaagggacg gatttgttgc
     1141 cattaaggga cggatttgtt gttaagaagg gacggatttg ttgttgtaaa gggacggatt
     1201 tgttgtattg tgggacgcag atacagtgtc cccttataca caaggaatgt cgaacgtggc
     1261 ctcaccccca atggtttaca aaagcaatgc cctggtcgag gccgcgtatc gcctcagtgt
     1321 tcaggaacag cggatcgttc tggcctgtat tagccaggtg aagaggagcg agcctgtcac
     1381 cgatgaagtg atgtattcag tgacggcgga ggacatagcg acgatggcgg gtgtccctat
     1441 cgaatcttcc tacaaccagc tcaaagaagc ggccctgcgc ctgaaacggc gggaagtccg
     1501 gttaacccaa gagcccaatg gcaaggggaa aagaccgagt gtgatgatta ccggctgggt
     1561 gcaaacaatc atctaccggg agggtgaggg ccgtgtagaa ctcaggttca ccaaagacat
     1621 gctgccgtac ctgacggaac tcaccaaaca gttcaccaaa tacgccttgg ctgacgtggc
     1681 caagatggac agcacccacg cgatcaggct ttacgagctg ctcatgcaat gggacagcat
     1741 cggccagcgc gaaatagaaa ttgaccagct gcgaaagtgg tttcaactgg aaggccggta
     1801 tccctcgatc aaggacttca agttgcgagt gcttgatcca gccgtgacgc agatcaacga
     1861 gcacagcccg ctacaggtgg agtgggcgca gcgaaagacc gggcgcaagg tcacacatct
     1921 gttgttcagt tttggaccga agaagcccgc caaggcggtg ggtaaggccc cagcgaagcg
     1981 caaggccggg aagatttcag atgctgagat cgcgaaacag gctcgccctg gtgagacatg
     2041 ggaagcggcc cgcgctcgac taacccagat gccgctggat ctggcctaga ggtgcaccac
     2101 ctgctttcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     2161 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     2221 cccagtcacg acgttgtaaa acgacggcca gtgccaagct tattatgaca acttgacggc
     2281 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
     2341 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
     2401 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
     2461 gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg
     2521 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
     2581 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
     2641 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
     2701 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
     2761 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
     2821 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
     2881 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
     2941 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
     3001 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
     3061 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
     3121 atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag aaaccaattg
     3181 tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccaa
     3241 accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa
     3301 aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg
     3361 cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg
     3421 ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggaat tcgagctcta
     3481 aggaggttat aaaaaatgga tattaatact gaaactgaga tcaagcaaaa gcattcacta
     3541 accccctttc ctgttttcct aatcagcccg gcatttcgcg ggcgatattt tcacagctat
     3601 ttcaggagtt cagccatgaa cgcttattac attcaggatc gtcttgaggc tcagagctgg
     3661 gcgcgtcact accagcagct cgcccgtgaa gagaaagagg cagaactggc agacgacatg
     3721 gaaaaaggcc tgccccagca cctgtttgaa tcgctatgca tcgatcattt gcaacgccac
     3781 ggggccagca aaaaatccat tacccgtgcg tttgatgacg atgttgagtt tcaggagcgc
     3841 atggcagaac acatccggta catggttgaa accattgctc accaccaggt tgatattgat
     3901 tcagaggtat aaaacgaatg agtactgcac tcgcaacgct ggctgggaag ctggctgaac
     3961 gtgtcggcat ggattctgtc gacccacagg aactgatcac cactcttcgc cagacggcat
     4021 ttaaaggtga tgccagcgat gcgcagttca tcgcattact gatcgttgcc aaccagtacg
     4081 gccttaatcc gtggacgaaa gaaatttacg cctttcctga taagcagaat ggcatcgttc
     4141 cggtggtggg cgttgatggc tggtcccgca tcatcaatga aaaccagcag tttgatggca
     4201 tggactttga gcaggacaat gaatcctgta catgccggat ttaccgcaag gaccgtaatc
     4261 atccgatctg cgttaccgaa tggatggatg aatgccgccg cgaaccattc aaaactcgcg
     4321 aaggcagaga aatcacgggg ccgtggcagt cgcatcccaa acggatgtta cgtcataaag
     4381 ccatgattca gtgtgcccgt ctggccttcg gatttgctgg tatctatgac aaggatgaag
     4441 ccgagcgcat tgtcgaaaat actgcataca ctgcagaacg tcagccggaa cgcgacatca
     4501 ctccggttaa cgatgaaacc atgcaggaga ttaacactct gctgatcgcc ctggataaaa
     4561 catgggatga cgacttattg ccgctctgtt cccagatatt tcgccgcgac attcgtgcat
     4621 cgtcagaact gacacaggcc gaagcagtaa aagctcttgg attcctgaaa cagaaagccg
     4681 cagagcagaa ggtggcagca tgacaccgga cattatcctg cagcgtaccg ggatcgatgt
     4741 gagagctgtc gaacaggggg atgatgcgtg gcacaaatta cggctcggcg tcatcaccgc
     4801 ttcagaagtt cacaacgtga tagcaaaacc ccgctccgga aagaagtggc ctgacatgaa
     4861 aatgtcctac ttccacaccc tgcttgctga ggtttgcacc ggtgtggctc cggaagttaa
     4921 cgctaaagca ctggcctggg gaaaacagta cgagaacgac gccagaaccc tgtttgaatt
     4981 cacttccggc gtgaatgtta ctgaatcccc gatcatctat cgcgacgaaa gtatgcgtac
     5041 cgcctgctct cccgatggtt tatgcagtga cggcaacggc cttgaactga aatgcccgtt
     5101 tacctcccgg gatttcatga agttccggct cggtggtttc gaggccataa agtcagctta
     5161 catggcccag gtgcagtaca gcatgtgggt gacgcgaaaa aatgcctggt actttgccaa
     5221 ctatgacccg cgtatgaagc gtgaaggcct gcattatgtc gtgattgagc gggatgaaaa
     5281 gtacatggcg agttttgacg agatcgtgcc ggagttcatc gaaaaaatgg acgaggcact
     5341 ggctgaaatt ggttttgtat ttggggagca atggcgatga cgcatcctca cgataatatc
     5401 cgggtaggcg caatcacttt cgtctactcc gttacaaagc gaggctgggt atttcccggc
     5461 ctttctgtta tccgaaatcc actgaaagca cagcggctgg ctgaggagat aaataataaa
     5521 cgaggggctg tatgcacaaa gcatcttctg ttgagttaag aacgagtatc gagatggcac
     5581 atagccttgc tcaaattgga atcaggtttg tgccaatacc agtagtctag aggatccccg
     5641 ggtaccgagc tcgaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5701 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5761 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5821 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5881 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5941 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6001 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6781 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6841 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6901 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6961 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7021 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7081 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7141 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7201 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7261 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7321 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7381 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7441 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7501 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7561 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7621 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7681 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7741 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7801 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     7861 aaataggcgt atcacgaggc cctttcgtc
//
