LOCUS       40924_3401        9602 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pABC1-loxB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9602)
  AUTHORS   Doehlemann J, Wagner M.
  TITLE     A family of single copy repABC-type shuttle vectors stably 
            maintained in the alpha-proteobacterium Sinorhizobium meliloti
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9602)
  AUTHORS   Doehlemann J, Wagner M.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-OCT-2016) LOEWE-Zentrum fuer Synthetische 
            Mikrobiologie, Philipps-Universitaet Marburg, Hans-Meerwein-Strasse,
            Marburg, Hesse 35043, Germany
REFERENCE   3  (bases 1 to 9602)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9602)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-OCT-2016) LOEWE-Zentrum fuer Synthetische Mikrobiologie, 
            Philipps-Universitaet Marburg, Hans-Meerwein-Strasse, Marburg, Hesse
            35043, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..4558
                     /label=oriVSm part (oriV_p42b) derived from pLoriV-42b
                     /note="oriVSm part (oriV_p42b) derived from pLoriV-42b"
     terminator      4614..4641
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      4779..4806
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      4943..5029
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    5113..5142
                     /label=MCS
                     /note="MCS"
     protein_bind    complement(5143..5176)
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      5208..5235
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      5328..5916
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6058..6503
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             6504..7922
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     oriT            complement(8279..8388)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    complement(8531..8564)
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(8590..8617)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(8708..9496)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gaaactgtca ctggtccgtt gatacaatcc agtcgtcaaa aaggcttttc acgtcaactc
       61 caaccgcacc gtttctccag cgaacgccgg ccgatcgtcg atcaggtcct tccctcgcct
      121 tgccaatcaa aaatcttgta acttcatgat ctcaaaggaa gaatatccta acaagaagtt
      181 atcaccctgt ttatttccgt cacctttcgt taaccataga tcgccacgga cagatgacac
      241 gaaatttttc atccggtctt gcctctgatt ggcgaattgg tgcggacaag cgccatcaac
      301 atcggtccga tggcaaattc gcctctctcg gcgcggcggg tcaggtcacg gaggtaacca
      361 ccggcggaat tgatgtgccc tgccctttca agcacgcacg ctatgacggt agccgcattt
      421 tcaggcccca tgacctcgca ggcctgttca taggccgagg ggctcactcc gagcattgag
      481 cgcacgacga cagccgccgc catcagatcg cgccatgtgc ccaccgagcc ctgcggtcca
      541 taggcggcga tctctgggca ggcctgcagc accaggccca gcgggaagga tttgagcccg
      601 ccgccggcaa ccgaggctga gtttgcagct ggcatgggcc tgcccttccc ctcctgtgtc
      661 gctattctgg ccctcggctc ggcccaacct tccgattggt cgtccaccgt tgcgccctgc
      721 ttcgtttcga agcgtggttc aagttcagta gtgtagtgtg ggtttgaatt ctgtatgtgc
      781 cgctcagaat gttgggcatt gccgctttga tttggagctt taacctgcat ttccaactga
      841 ttggtgattt cttcgcgcaa cagttccagt tcgtcgagtg cggcggcaat cttctcgaag
      901 gatggcgagc gcggcagccg ttcgacgatg cctcgaaaat gcaggtggac cccgctccaa
      961 tcaccagagg cgccctcttc cacggccatc tcgatgagct tggcgatgtc gcgcctgcaa
     1021 agcgtcaggc gctctttgag ccgctgcagc tgcaatcgtt cggctgccat ctcggccgcc
     1081 agccgctcga tctcgtccgc gcgcgcaagc agcggcgcca gcgagaagcc gaatgcctca
     1141 cggatctccc cgccctgctc cttgcgagcg taacgcttgc cgttcgggct gtccttgcgg
     1201 atcagcaatc cggcctcgat cagcgccgcc aggtggcgac ggatcgtctg ctccgccatt
     1261 ccatgcgcgc gcagagagag ctgcgtattc gaaggaaata cgaccaagcc gttctcctcg
     1321 gagagctctc ccttgggata aaagctcagc aaggcgttca ggatggaaag cgcccggtct
     1381 gtaatgccga gcagcggcct ggcctcgcac agggcgcgat agagtttcca tttatcaatc
     1441 gatcggccgg gctcgatttt tcgggcagcg gcctgacttg ccaacatgcc aaacgtcatc
     1501 ggccgccgcc cgaagggcgt cgtcacactt ccgctttcca ttttcttcgc cttcttcaag
     1561 gcaaaagatc acgcctcacc aaattcggtg ccaaagactc ttgactatga ttcgcggaaa
     1621 tgtgattctc tgggtgtcta gatcagagag ggcttccgcg acggcaacgt tcgggggcct
     1681 tttcttttgc ttattgatct ccgttcttcg tcaaatcctg tgtcttctcg aacgcctcgt
     1741 agagacgatc cagattgctg gcgatgtagg cgccaaaggc agatgccttc tccgccttca
     1801 gcgcgatggt gaactgtttt ccatcatcct tgatcttcgc tttgacggct ccgtcccggc
     1861 gctgccaggc gtgcgccacg ggctgaactg cggcggcggg cgacgcctgt tggttcctgc
     1921 tgatgaaggc gacaagcatg tcgaagcggg cgtccggctc ggccgtctcg aactccaccg
     1981 atctggtgaa ctcaatcgcc cgggctgcga tgttttccgt ctcgaatttc gccgaaagat
     2041 catgccagcg atcgcggcca gtcgtcttgg ctacgccgat cgcggccagg acttcggccg
     2101 gaatccgctt cgtgaccgac agcatcttgg acaccgtcgt cttgtctgcg ctgagtgccg
     2161 acatgatcgt ttcccggtcg aaacccaacg tgtcgagctt gtcggcaaac atcgtcctct
     2221 cgatgaagga gagatcggcg cgtgccgaat tctcctggcc ctgcgcaatg acatggtcgc
     2281 ggtcgtcgag tctcttgacc accgcccgga caggtcggcc gagctccttt gccgcccgcg
     2341 cacgacgatg accaaaagcg atctgatatc gcccctccct ctcgggatgc gggcgcacaa
     2401 ggatcggtga atcctggccg cgcaggcgga tcgcctcgac taattcccgg aactgttcgt
     2461 cagtatgcgc cagccggtcg gtcacgaagg aatcttcgat cagcgccggg tcgagatcga
     2521 tcaccgtctc gccgctcgcc agctgctctt cgatcgcctt ggcggcatcg gccttggcgg
     2581 ccaaggcatc gatgctgcgg gtgaccgcac cgagcgcgcc aatcccctta taggtgatat
     2641 gctgcctctc gtcgcgaggg gccggctcat cctcattgtt taccgcagta aacttcttgg
     2701 aatcgtccat caggcccgac agaagatttt tacgcgccat caaccagctc cttgccccgg
     2761 tttaccgcag taaacttccc gctcatttcc ggccccatgc cttatggatg agatcgacga
     2821 tttccgtgtt gactgactcc atcgcctcca tggcgcggtc ataggtcgcg cgggtaaact
     2881 gacttcgctc gacttcgtac agcgtctggt tagtcatcgc cgcatcggag atcgcgacgc
     2941 tgcgcagcat cgggttggtg agcacatggc tcttgaacat cgagcgcatg aaggcgacca
     3001 tctgcgtctg cggcccgtcc tgcggatcgt agcgggtcac gaggtaacgc agccagtcga
     3061 ggttcatatt gccgccggct cccttcagcg tgttcagcac ttcgccaagc atttgcagga
     3121 actggcacat cgacatgaca tcgagcatct gcggatgaac ggtgatcaga accgccgtcg
     3181 caccgcaaat ggcgctcatg gtcaaaaagc caagctgggg agggcaatcg atgatgacga
     3241 cgtcgtaatc atccgcgacc gaggccagcg cctcatccag ccgggcaaag aagacgcgcc
     3301 cataatcgct cgctttgccc tgagcgagga cccgcggcgt atcgtgctcg aattccatca
     3361 gttcgagatt gcccggcacg agatgaaggt tcgggaagtt ggtcggccgg ataatgtccc
     3421 gcagcggacg ccgctgatcg tcatagcgaa tggcggcgta gagtgtctcg ttttcgttga
     3481 cgtcgaactc cggctgaaag ccgtgtatgg cagaaagaga cgcttgcgga tcgagatcga
     3541 cggccaggac gcggtgaccg gtcagggcga gatgctgggc aagatgcgcg gcactcgtcg
     3601 tcttgccgct gccccccttg aagttgacga cagcaacgac ctgcagatgt tcgtggccgc
     3661 ggcgatgacg cacataatgt gttccggcgc gcgcattatg ctcgagaaag ccacgcatct
     3721 cctccatctg ctccgccgta tacgatcggc ggccagacgg cgtaacctgg gggagggcac
     3781 ccttgccctc gagcgaaaga tttcgaagat agccgctggt cacgccgaga aaccgcgcca
     3841 cttcagcaag ctgaaattcc cgcagaccct ttagcgcgcg cggaggaaac atttcgaggc
     3901 gatgctgctt cagcttgtcg gatagttcct gtgcctgacc gatgatcagc tggtcgacat
     3961 cggaaattgc tttttctatt tgcggtgcca tattcatgga aatctcgaaa atgcgattta
     4021 aaagctctac gcgctttgaa accgcattta gcgccgattc tggaggggtg gcaaggcaaa
     4081 tagggttaac aagatcctat caaaaggttg tccagtctat ttcaagcgct tgcgccggcc
     4141 gggcgtgccg ctggcgggtc cagcaatcgc acattcacag gccgaatcgg acgagaccgg
     4201 ccgggaacag cagcagtata atccgaatcg caatgtcggg cgtgcaaagc gccttgctgc
     4261 aagcccgcag atgcaaggca tcggttcgct ggcgtggatg gagcggggcg cggcggtaag
     4321 gtgccgggtc tggtcgatca gccgtaactt gccagatcat tcgagcctga gttcacaggg
     4381 gaattgccgc ctggccagct actccagagg cggacgggtg atattcgcta tcgaaggcaa
     4441 agccggaccg ctttccgcct gcgccatcgc ttttaccaat gcgatgatcg tttcacgaat
     4501 atcgggatcg gcaatcttca gaaagactaa cttctcgtgg aactctatcg taactgaagt
     4561 ctattgaagg aacactgtat ctcggtcaag tgggaagagt ttgtagaaac gcaaaaaggc
     4621 catccgtcag gatggccttc tgcttaattt gatggtcgac tcggcggaaa gcgagaagag
     4681 gttcgcctcg gtctcgccgg cggcgaatcc ggccgtcggg ccggtcggcc ccttccggaa
     4741 gcgaacgcga accgtcttca agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     4801 ccttctgctt aatttgatgg aactcggagc gtccgcgtcc tcgtaaacga gggcgagcgg
     4861 ccggttaacg gtcgcggagt ccgaacggtt cttccgagtt cgaatccgcg ggtctccggg
     4921 agccccgaga taaccggcgc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     4981 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgcgatcccg
     5041 tagccgatcg cgccgaagac ctccgcgccg ccggagcccg aactctcggc gaggaagccg
     5101 accgtcgtcg gggtttaaac ttaattaagc ggccgcagta cttaccgttc gtatagcata
     5161 cattatacga agttatcata tgcccactaa gagtttgtag aaacgcaaaa aggccatccg
     5221 tcaggatggc cttctgctta atttgatggt tatcagaaga gtgtaagcgg gttgactata
     5281 ctactgtttc agactggcgt aatcactcag tagatcaaag gatcttcttg agatcctttt
     5341 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5401 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5461 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5521 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5581 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5641 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5701 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5761 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5821 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5881 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc gactggcaat
     5941 cactgagtaa gactattctg tccatgtgac gataagttcc ctactgacag aatcagtgac
     6001 gagttcttct gagcgggact ctggggttcg ctagaggatc gatccttttt aacccatcac
     6061 atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt tctgaattgt
     6121 gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca gagaatgaaa
     6181 agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt ttatgatttt
     6241 ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct aatagaatga
     6301 ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc taaaatgtgt
     6361 aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt tttttattgt
     6421 gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg ctaacacagt
     6481 acataaaaaa ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa gcaacagtat
     6541 taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg aaagaaacga
     6601 accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat gatatgctgc
     6661 aaatccctga acagcaaaaa aatgaaaaat atcaagtttc tgaatttgat tcgtccacaa
     6721 ttaaaaatat ctcttctgca aaaggcctgg acgtttggga cagctggcca ttacaaaacg
     6781 ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta gccggagatc
     6841 ctaaaaatgc ggatgacaca tcgatttaca tgttctatca aaaagtcggc gaaacttcta
     6901 ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc gatgcaaatg
     6961 attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt acatctgacg
     7021 gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa caaacactga
     7081 caactgcaca agttaacgta tcagcatcag acagctcttt gaacatcaac ggtgtagagg
     7141 attataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtacag cagttcatcg
     7201 atgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac tacgtagaag
     7261 ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat ggctaccaag
     7321 gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc ttccgtcaag
     7381 aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca aacggcgctc
     7441 tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa ccgctgattg
     7501 catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg aacggcaaat
     7561 ggtacctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt acgtctaacg
     7621 atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag ccgctgaaca
     7681 aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt acttactcac
     7741 acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat atgacaaaca
     7801 gaggattcta cgcagacaaa caatcaacgt ttgcgccgag cttcctgctg aacatcaaag
     7861 gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta acagttaaca
     7921 aataaaaacg caaaagaaaa tgccgatggg taccgagcga aatgaccgac caagcgacgc
     7981 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     8041 gaatcgtttt ccgggacgcc ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg
     8101 tagccctggc cgacggccag caggtaggcc gacaggctca tgccggccgc cgccgccttt
     8161 tcctcaatcg ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc
     8221 ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc
     8281 ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg
     8341 aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt
     8401 tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa
     8461 aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa
     8521 aagccatatg ataacttcgt atagcataca ttatacgaac ggtagtcgac aagagtttgt
     8581 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct taatttgatg actggttaca
     8641 gtgtcacccg ttatcaatag agggaccttt tctccgacga atagagtaac agaagtcggg
     8701 atccttattt gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca
     8761 actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt
     8821 caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat
     8881 ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta
     8941 catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta
     9001 gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta
     9061 ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg
     9121 tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt
     9181 cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta
     9241 cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca
     9301 aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac
     9361 tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg
     9421 gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga
     9481 tcaccgcttc cctcatatgt tttctcctct gctacgtaac gcgtctactt gcgtactagt
     9541 taactagttc gatgattaat tgtcaacaga ctgattggtc aggcacaact ttcagataag
     9601 ac
//
