LOCUS       40924_994        6643 bp DNA     linear   UNA 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6643)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6643
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(1..104)
                     /label=AmpR promoter
     rep_origin      complement(130..585)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     727..743
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1241..1265
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1597..2388
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(2711..2727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2735..2751)
                     /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(2759..2789)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2804..2825)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3058..3503
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3504..4922
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     rep_origin      complement(5023..5611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5785..6642)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
       61 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
      121 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
      181 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      241 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
      301 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
      361 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
      421 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
      481 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
      541 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
      601 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      661 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      721 gacgttgtaa aacgacggcc agtgagcgcg ttccaactcg cttaattgcg agtttttatt
      781 tcgtttattt caattaaggt aactaaaaaa ctccttatac ctcgaggaaa cgccggcgaa
      841 agcggccgca aacgtcgacg aaagcgatcg caaattaatt aaaaacgcgc gcgaaacctc
      901 agcaaagacg tcaaaaccgg taaacctagg aaaactagta aaaagcttag agctagcaaa
      961 caattgaaag ggcccaaaaa cgttaaatct agaaggcctc ccgggaaagg atccaaagaa
     1021 ttcaaagtcg acaaaatcga taaagagctc aaacctgcag gaaaggtacc aaaccatgga
     1081 aaggcgcgcc tttataacgg tcctaaggta gcgaaaaata gggataacag ggtaatttta
     1141 cgcgtaaatg tttagttcct cgttttctct cgttggaaga agaagaaacg agaaactaaa
     1201 attaaaaagt acttttagat ctgtttaaac tatcagtgtt tgacaggata tattggcggg
     1261 taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg gcgtgaaaag
     1321 gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccct cgggagaaac
     1381 gtacgctcgc gttggcagca tcacccataa ttgtggtttc aaaatcggct ccgtcgatac
     1441 tatgttatac gccaactttg aaaacaactt tgaaaaagct gttttctggt atttaaagtt
     1501 ttagaatgca aggaacagtg aattggagtt cgtcttgtta taattagctt cttggggtat
     1561 ctttaaatac tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag aatatcaccg
     1621 gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct
     1681 gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc
     1741 cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa
     1801 ggaaagctgc ctgttccaaa ggtcctgcac cttgaacggc atgatggctg gagcaatctg
     1861 ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct
     1921 gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg
     1981 gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat
     2041 aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag acactccatt taaagatccg
     2101 cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac
     2161 ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat
     2221 cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgatc
     2281 agggaggata ttggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag
     2341 cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta gtacctagat
     2401 gtggcgcaac gatgccggcg acaagcagga gcgcaccgac ttcttccgca tcaagtgttt
     2461 tggctctcag gccgaggccc acggcaagta tttgggcaag gggtcgctgg tattcgtgca
     2521 gggcaagatt cggaatacca agtacgagaa ggacggccag acggtctacg ggacgtacga
     2581 aaggtcaaat tgggtatacc catttgggcc tagtctagcc ggcatggcgc attacagcaa
     2641 tacgcaattt aaatgcgcct agcgcatttt cccgacctta atgcgcctaa acgcgcttgg
     2701 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2761 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2821 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2881 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2941 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatggcac
     3061 atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt tctgaattgt
     3121 gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca gagaatgaaa
     3181 agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt ttatgatttt
     3241 ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct aatagaatga
     3301 ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc taaaatgtgt
     3361 aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt tttttattgt
     3421 gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg ctaacacagt
     3481 acataaaaaa ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa gcaacagtat
     3541 taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg aaagaaacga
     3601 accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat gatatgctgc
     3661 aaatccctga acagcaaaaa aatgaaaaat atcaagttcc tgagttcgat tcgtccacaa
     3721 ttaaaaatat ctcttctgca aaaggtctgg acgtttggga cagctggcca ttacaaaacg
     3781 ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta gccggagatc
     3841 ctaaaaatgc ggatgacacc tcgatttata tgttctatca aaaagtcggc gaaacttcta
     3901 ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc gatgcaaatg
     3961 attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt acatctgacg
     4021 gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa caaacactga
     4081 caactgcaca agtcaacgta tcagcatcag acagctcttt gaacatcaac ggtgtagagg
     4141 actataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtccag cagttcatcg
     4201 acgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac tacgtagaag
     4261 ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat ggctaccaag
     4321 gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc ttccgtcaag
     4381 aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca aacggcgctc
     4441 tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa ccgctgattg
     4501 catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg aacggcaaat
     4561 ggtatctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt acgtctaacg
     4621 atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag ccgctgaaca
     4681 aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt acttactcac
     4741 acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat atgacaaaca
     4801 gaggattcta cgcagacaaa caatcaacgt ttgcgccgag cttcctgctg aacatcaaag
     4861 gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta acagtcaaca
     4921 aataaaaacg caaaagaaaa tgccgatatg ctatcggcat tacatgtgag caaaaggcca
     4981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5461 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc ata
//
