LOCUS       sequence-274478-        6640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  matBC cassette from R. trifolii controlled by a Plac promoter; 
            pSC101 ori, cat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  AUTHORS   Jeschek M, Bahls MO, Schneider V, Marliere P, Ward TR, Panke S
  TITLE     Biotin-independent strains of Escherichia coli for enhanced 
            streptavidin production.
  JOURNAL   Metab Eng. 2017 Mar;40:33-40. doi: 10.1016/j.ymben.2016.12.013. Epub
            2017 Jan 3.
  PUBMED    28062280
REFERENCE   2  (bases 1 to 6640)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2017-03"; volume: "40:33-40. doi"; pages: " 
            10.1016/j.ymben.2016.12.013. Epub 2017 Jan 3"
COMMENT     Sequence Label: pCKmatBC
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      3158..3229
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3245..3272
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3328..3345)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3328..3344)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3337..3359)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3788..4447)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4362..4381
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4448..4550)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4895..5117
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5165..6115
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgaattg cggccgcgcg aattcgagct cggtacccgg ggatccagga gggcgaaagt
      181 gagcaaccat cttttcgacg ccatgcgggc cgccgcgccc ggtaacgcac cattcatccg
      241 gatcgataac acgcgcacat ggacctatga cgacgccttc gctctttccg gccgcattgc
      301 cagcgcgatg gacgcgctcg gcattcgccc cggcgaccgc gttgcggtgc aggtcgagaa
      361 aagtgccgag gcattgatcc tctatctcgc ctgtcttcga agcggcgccg tctacctgcc
      421 gctcaacacc gcctatacgc tggctgagct cgattatttt atcggcgatg cggagccgcg
      481 tttggtggtt gtcgcatcgt cggctcgagc gggcgtggag acaatcgcca agccccgcgg
      541 tgcgatcgtc gaaactctcg acgctgctgg cagcggctcg ttgctggatc tcgcccgcga
      601 cgagccggcc gactttgtcg atgcctcgcg ctccgccgat gatctggcgg cgatcctcta
      661 cacgtccgga acgacgggac gctccaaggg ggcgatgctc acgcatggga acctgctctc
      721 gaacgccctg accttgcgag atttttggcg cgtcaccgcc ggcgatcgac tgatccatgc
      781 cttgccgatc ttccacacgc atggactgtt cgtcgccacg aacgtcacac tgctcgccgg
      841 cgcctcgatg ttcctgctgt cgaagttcga cccggaggag atcctgtcgc tgatgccgca
      901 ggcaacgatg ctgatgggcg tgccgacctt ctacgtgcgc ctcctgcaga gcccgcgcct
      961 cgacaagcaa gcggtcgcca acatccgcct cttcatttcc ggttcggctc cactgcttgc
     1021 agaaacacat accgagttcc aggcacgtac cggtcacgcc attctcgagc gctacggcat
     1081 gacggaaacc aatatgaaca cgtccaaccc ttatgagggg aaacggattg ccggaacggt
     1141 cggcttcccg ctgcctgatg tgacggtgcg cgtcaccgat cccgccaccg ggctcgcgct
     1201 gccgcccgaa caaaccggca tgatcgagat caaggggccg aacgttttca agggctattg
     1261 gcgcatgccc gaaaaaaccg cggccgaatt caccgccgac ggtttcttca tcagcggcga
     1321 tctcggcaag atcgaccgcg acggttatgt ccacatcgtc ggccgcggca aggatctggt
     1381 gatttcgggt ggatacaaca tctatccgaa agaggttgag ggcgagatcg accagatcga
     1441 gggtgtggtt gagagcgctg tgatcggcgt gccgcatccc gatttcggag aaggcgtaac
     1501 ggccgtcgtc gtgcgcaagc ccggcgctgc cctcgatgaa aaggccatcg tcagcgccct
     1561 ccaggaccgg ctcgcgcgct acaaacaacc caagcgcatc atctttgcag aggacttgcc
     1621 gcgcaacacg atgggtaagg ttcagaaaaa catcctgcgg cagcaatacg ccgatcttta
     1681 taccaggacg taaggcgacc gcgctctctg ggaggagagt gcgtcgacat cccgcatcaa
     1741 tcttgaaaac agcaactgcg acgcggaggc gtcggaggga ggggaatcat gggtattgaa
     1801 ttactgtcca taggcctgct gatcgccatg ttcatcattg cgacgatcca gccaatcaac
     1861 atgggtgcgc tcgcctttgc cggcgccttc gtgctcggct cgatgatcat cgggatgaaa
     1921 accaacgaaa tatttgccgg ctttccgagt gatctgttcc tgacgctcgt cgccgtcacc
     1981 tacctcttcg ccatagcgca gatcaacggc acgatcgact ggctcgtcga atgtgccgtc
     2041 cgcctggtac gcgggcggat cggcttgatt ccctgggtga tgttccttgt cgccgccatc
     2101 attactggct tcggtgcact tgggcctgct gcggtcgcca ttctcgcacc cgtcgcgttg
     2161 agctttgccg tgcagtaccg cattcatccg gtgatgatgg gtctgatggt gatccacggc
     2221 gcgcaggcag gcggcttctc gccgatcagc atctatggcg gaatcaccaa ccagatcgtt
     2281 gcgaaggccg gcctgccttt cgctccgacc tcgctgtttc tttccagctt cttctttaac
     2341 ctggcgatcg cggtgctggt gttcttcgtg ttcggcggcg cgagggtgat gaagcacgat
     2401 cccgcatcac ttggcccctt gcccgaactc catcccgagg gcgtatcggc gtcgatcaga
     2461 ggccacggcg gcacgccggc aaaaccgatc agagagcatg cctatggtac ggcggccgat
     2521 accgcgacga cgttgcgtct gaacaatgag agaattacca ccttgatcgg cctgacggcg
     2581 ctcggcatcg gcgccctggt tttcaagttc aatgttggcc tcgtcgccat gaccgtcgcc
     2641 gtcgtcctcg cgctgctgtc accgaagacc cagaaggccg caatcgacaa ggtcagttgg
     2701 tcgaccgtgc tgctgattgc cggcatcatc acctatgtcg gcgtcatgga gaaggccggt
     2761 acggtcgact acgtggcgaa tggcatatcc agtctcggca tgccgctact ggtagcgctc
     2821 ctgctttgct ttacgggcgc catcgtctcg gcctttgctt cctcgaccgc gctgctcggc
     2881 gcgatcatcc cgcttgccgt tccattcctc ctgcaagggc acatcagcgc catcggtgtg
     2941 gtcgcggcga tcgccatctc gacgacgatc gtcgacacca gcccattctc caccaacggc
     3001 gcccttgtcg tcgccaatgc gccggacgac agccgtgagc aggtgttgcg acagctactg
     3061 atctacagcg ccttgatcgc tatcatcggt ccgatcgttg cctggttggt gttcgtcgtg
     3121 cccgggctgg tttgaggcgc gccgatgagc caggcatcaa ataaaacgaa aggctcagtc
     3181 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     3241 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagtcga cctgcaggca
     3301 tgcaagcttg cggccgccca gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     3361 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3421 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3481 atggcgctaa ccgtttttat caggctctgg gaggcagaat aaatgatcat atcgtcaatt
     3541 attacctcca cggggagagc ctgagcaaac tggcctcagg catttgagaa gcacacggtc
     3601 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     3661 cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc
     3721 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
     3781 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     3841 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     3901 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     3961 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     4021 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     4081 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     4141 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     4201 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     4261 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     4321 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     4381 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     4441 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     4501 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
     4561 ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt
     4621 ctgcgaagtg atcttccgtc acaggtattt attcggcgcc tgtagtgcca tttaccccca
     4681 ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg
     4741 tgcctgatgg tcggagacaa aaggaatatt cagcgatttg cccgagcttg cgagggtgct
     4801 acttaagcct ttagggtttt aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat
     4861 ccttttgtaa tactgcggaa ctgactaaag tagtgagtta tacacagggc tgggatctat
     4921 tctttttatc tttttttatt ctttctttat tctataaatt ataaccactt gaatataaac
     4981 aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt ctagcagaat ttacaagttt
     5041 tccagcaaag gtctagcaga atttacagat acccacaact caaaggaaaa ggactagtaa
     5101 ttatcattga ctagcccatc tcaattggta tagtgattaa aatcacctag accaattgag
     5161 atgtatgtct gaattagttg ttttcaaagc aaatgaacta gcgattagtc gctatgactt
     5221 aacggagcat gaaaccaagc taattttatg ctgtgtggca ctactcaacc ccacgattga
     5281 aaaccctaca aggaaagaac ggacggtatc gttcacttat aaccaatacg ctcagatgat
     5341 gaacatcagt agggaaaatg cttatggtgt attagctaaa gcaaccagag agctgatgac
     5401 gagaactgtg gaaatcagga atcctttggt taaaggcttt gagattttcc agtggacaaa
     5461 ctatgccaag ttctcaagcg aaaaattaga attagttttt agtgaagaga tattgcctta
     5521 tcttttccag ttaaaaaaat tcataaaata taatctggaa catgttaagt cttttgaaaa
     5581 caaatactct atgaggattt atgagtggtt attaaaagaa ctaacacaaa agaaaactca
     5641 caaggcaaat atagagatta gccttgatga atttaagttc atgttaatgc ttgaaaataa
     5701 ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa ccaataagta aagatttaaa
     5761 cacttacagc aatatgaaat tggtggttga taagcgaggc cgcccgactg atacgttgat
     5821 tttccaagtt gaactagata gacaaatgga tctcgtaacc gaacttgaga acaaccagat
     5881 aaaaatgaat ggtgacaaaa taccaacaac cattacatca gattcctacc tacataacgg
     5941 actaagaaaa acactacacg atgctttaac tgcaaaaatt cagctcacca gttttgaggc
     6001 aaaatttttg agtgacatgc aaagtaagta tgatctcaat ggttcgttct catggctcac
     6061 gcaaaaacaa cgaaccacac tagagaacat actggctaaa tacggaagga tctgaggttc
     6121 ttatggctct tgtatctatc agtgaagcat caagactaac aaacaaaagt agaacaactg
     6181 ttcaccgtta catatcaaag ggaaaactgt ccatatgcac agatgaaaac ggtgtaaaaa
     6241 agatagatac atcagagctt ttacgagttt ttggtgcatt taaagctgtt caccatgaac
     6301 agatcgacaa tgtaacagat gaacagcatg taacacctaa tagaacaggt gaaaccagta
     6361 aaacaaagca actagaacat gaaattgaac acctgagaca acttgttaca gctcaacagt
     6421 cacacataga cagcctgaaa caggcgatgc tgcttatcga atcaaagctg ccgacaacac
     6481 gggagccagt gacgcctccc gtggggaaaa aatcatggca attctggaag aaatagcgcc
     6541 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6601 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
//
