LOCUS       sequence-264187-        8597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre recombinase for excising chromosomal loxP-Hyg-loxP 
            cassette inserts..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8597)
  AUTHORS   Murphy KC, Papavinasasundaram K, Sassetti CM
  TITLE     Mycobacterial recombineering.
  JOURNAL   Methods Mol Biol. 2015;1285:177-99. doi: 
            10.1007/978-1-4939-2450-9_10.
  PUBMED    25779316
REFERENCE   2  (bases 1 to 8597)
  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: "Methods Mol
            Biol."; date: "2015"; volume: "1285:177-99. doi"; pages: " 
            10.1007/978-1-4939-2450-9_10"
COMMENT     Sequence Label: pCre-SacB-zeo
FEATURES             Location/Qualifiers
     source          1..8597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             20..1051
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     terminator      1093..1139
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1289..1308)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1373..1392)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     polyA_signal    1992..2126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2016..2035)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2070..2089
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2190..2564)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3172..3760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3661..3680
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6271..6716
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(6717..6803,6804..8138)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 ggcgccagag gcactcgaga tgtccaattt actgaccgta caccaaaatt tgcctgcatt
       61 accggtcgat gcaacgagtg atgaggttcg caagaacctg atggacatgt tcagggatcg
      121 ccaggcgttt tctgagcata cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc
      181 atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta
      241 tcttctatat cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgggcca
      301 gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc
      361 actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc
      421 tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg
      481 ctgccaggat atacgtaatc tggcatttct ggggattgct tataacaccc tgttacgtat
      541 agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt
      601 aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag
      661 cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc
      721 gaataactac ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag
      781 ccagctatca actcgcgccc tggaagggat ttttgaagca actcatcgat tgatttacgg
      841 cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg
      901 agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg
      961 ctggaccaat gtaaatattg tcatgaacta tatccgtaac ctggatagtg aaacaggggc
     1021 aatggtgcgc ctgctggaag atggcgatta gcgtcgacgt agttaactag cgtacgatcg
     1081 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatg
     1141 ccatcatggc cgcggtgatc agctagccac ctgacgtcgg ggggggggga aagccacgtt
     1201 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1261 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1321 acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1381 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1441 catgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1501 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1561 atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc
     1621 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     1681 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1741 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     1801 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa tcttttgcca
     1861 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     1921 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     1981 gatcttgcca tgatccagac atgataagat acattgatga gtttggacaa accacaacta
     2041 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2101 ccattataag ctgcaataaa caagtttcga ggtcgacccc cctcggaccc gtcgggccgc
     2161 gtcggaccgg cggtgttggt cggcgtcggt cagtcctgct cctcggccac gaagtgcacg
     2221 cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg
     2281 gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg
     2341 tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg
     2401 tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc
     2461 tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg
     2521 tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatggtgaa tcctcctgaa
     2581 tatgtagagc ttctttcctg cactatgatg agtgcacccc gctcaagctt gcctcgacgg
     2641 cggtgtgctg tcaattcaga gctgagcctg atgcactcaa cttactgagc atgctaacgc
     2701 tggtcgtgcg ggtcggaatt atggaactgc ctcggtgagt tttctccttc attacagaaa
     2761 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2821 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2881 acgctgactt gacgggacgg cggctttgtt gaataaatcg aacttttgct gagttgaagg
     2941 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     3001 tcaccaactg gtccacctac aacaaagctc tcaccaaccg tggctccctc actttctggc
     3061 tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac
     3121 ctcactagtt ccatgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3181 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3241 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3301 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3361 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3421 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3481 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3541 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgagg ggagaaaggc
     3601 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3661 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3721 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3781 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3841 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3901 aacgaccgag cgcaacgcgt gagcccacca gctccgtaag ttcgggcgct gtgtggctcg
     3961 tacccgcgca ttcaggcggc agggggtcta acgggtctaa ggcggcgtgt acggccgcca
     4021 cagcggctct cagcggcccg gaaacgtcct cgaaacgacg catgtgttcc tcctggttgg
     4081 tacaggtggt tgggggtgct cggctgtcgc tggtgttcca ccaccagggc tcgacgggag
     4141 agcgggggag tgtgcagttg tggggtggcc cctcagcgaa atatctgact tggagctcgt
     4201 gtcggaccat acaccggtga ttaatcgtgg tctactacca agcgtgagcc acgtcgccga
     4261 cgaatttgag cagctctggc tgccgtactg gccgctggca agcgacgatc tgctcgaggg
     4321 gatctaccgc caaagccgcg cgtcggccct aggccgccgg tacatcgagg cgaacccaac
     4381 agcgctggca aacctgctgg tcgtggacgt agaccatcca gacgcagcgc tccgagcgct
     4441 cagcgcccgg gggtcccatc cgctgcccaa cgcgatcgtg ggcaatcgcg ccaacggcca
     4501 cgcacacgca gtgtgggcac tcaacgcccc tgttccacgc accgaatacg cgcggcgtaa
     4561 gccgctcgca tacatggcgg cgtgcgccga aggccttcgg cgcgccgtcg atggcgaccg
     4621 cagttactca ggcctcatga ccaaaaaccc cggccacatc gcctgggaaa cggaatggct
     4681 ccactcagat ctctacacac tcagccacat cgaggccgag ctcggcgcga acatgccacc
     4741 gccgcgctgg cgtcagcaga ccacgtacaa agcggctccg acgccgctag ggcggaattg
     4801 cgcactgttc gattccgtca ggttgtgggc ctatcgtccc gccctcatgc ggatctacct
     4861 gccgacccgg aacgtggacg gactcggccg cgcgatctat gccgagtgcc acgcgcgaaa
     4921 cgccgaattt ccgtgcaacg acgtgtgtcc cggaccgcta ccggacagcg aggtccgcgc
     4981 catcgccaac agcatttggc gttggatcac aaccaagtcg cgcatttggg cggacgggat
     5041 cgtggtctac gaggccacac tcagtgcgcg ccagtcggcc atctcgcgga agggcgcagc
     5101 agcgcgcacg gcggcgagca cagttgcgcg gcgcgcaaag tccgcgtcag ccatggaggc
     5161 attgctatga gcgacggcta cagcgacggc tacaaccggc agccgactgt ccgcaaaaag
     5221 cggcgcgtga ccgccgccga aggcgctcga atcaccggac tatccgaacg ccacgtcgtc
     5281 cggctcgtgg cgcaggaacg cagcgagtgg ctcgccgagc aggctgcacg ccgcgaacgc
     5341 atccgcgcct atcacgacga cgagggccac tcttggccgc aaacggccaa acatttcggg
     5401 ctgcatctgg acaccgttaa gcgactcggc tatcgggcga ggaaagagcg tgcggcagaa
     5461 caggaagcgg ctcaaaaggc ccacaacgaa gccgacaatc caccgctgtt ctaacgcaat
     5521 tggggagcgg gtgtcgcggg ggttccgtgg ggggttccgt tgcaacgggt cggacaggta
     5581 aaagtcctgg tagacgctag ttttctggtt tgggccatgc ctgtctcgtt gcgtgtttcg
     5641 ttgcgcccgt tttgaatacc agccagacga gacggggttc tacgaatctt ggtcgatacc
     5701 aagccatttc cgctgaatat cggggagctc accgccagaa tcggtggttg tggtgatgta
     5761 cgtggcgaac tccgttgtag tgcctgtggt ggcatccgtg gcgcggccgc ggtaccagat
     5821 ctttaaatct agacggtgac cacaacgcgc ccgctttgat cggggacgtc tgcggccgac
     5881 catttacggg tcttgttgtc gttggcggtc atgggccgaa catactcacc cggatcggag
     5941 ggccgaggac aaggtcgaac gaggggcatg acccggtgcg gggcttcttg cactcggcat
     6001 aggcgagtgc taagaataac gttggcactc gcgaccggtg agtgctaggt cgggacggtg
     6061 aggccaggcc cgtcgtcgca gcgagtggca gcgaggacaa cttgagccgt ccgtcgcggg
     6121 cactgcgccc ggccagcgta agtagcgggg ttgccgtcac ccggtgaccc ccgtttcatc
     6181 cccgatccgg aggaatcact tcgcaatggc caagacaatt gcggatccag ctgcaggtcg
     6241 actctagagg atcgatcctt tttaacccat cacatatacc tgccgttcac tattatttag
     6301 tgaaatgaga tattatgata ttttctgaat tgtgattaaa aaggcaactt tatgcccatg
     6361 caacagaaac tataaaaaat acagagaatg aaaagaaaca gatagatttt ttagttcttt
     6421 aggcccgtag tctgcaaatc cttttatgat tttctatcaa acaaaagagg aaaatagacc
     6481 agttgcaatc caaacgagag tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg
     6541 ttactgataa agcaggcaag acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc
     6601 accccttaca tattttaggt ctttttttat tgtgcgtaac taacttgcca tcttcaaaca
     6661 ggagggctgg aagaagcaga ccgctaacac agtacataaa aaaggagaca tgaacgatga
     6721 acatcaaaaa gtttgcaaaa caagcaacag tattaacctt tactaccgca ctgctggcag
     6781 gaggcgcaac tcaagcgttt gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg
     6841 gcatttccca tattacacgc catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa
     6901 aatatcaagt tcctgaattc gattcgtcca caattaaaaa tatctcttct gcaaaaggcc
     6961 tggacgtttg ggacagctgg ccattacaaa acgctgacgg cactgtcgca aactatcacg
     7021 gctaccacat cgtctttgca ttagccggag atcctaaaaa tgcggatgac acatcgattt
     7081 acatgttcta tcaaaaagtc ggcgaaactt ctattgacag ctggaaaaac gctggccgcg
     7141 tctttaaaga cagcgacaaa ttcgatgcaa atgattctat cctaaaagac caaacacaag
     7201 aatggtcagg ttcagccaca tttacatctg acggaaaaat ccgtttattc tacactgatt
     7261 tctccggtaa acattacggc aaacaaacac tgacaactgc acaagttaac gtatcagcat
     7321 cagacagctc tttgaacatc aacggtgtag aggattataa atcaatcttt gacggtgacg
     7381 gaaaaacgta tcaaaatgta cagcagttca tcgatgaagg caactacagc tcaggcgaca
     7441 accatacgct gagagatcct cactacgtag aagataaagg ccacaaatac ttagtatttg
     7501 aagcaaacac tggaactgaa gatggctacc aaggcgaaga atctttattt aacaaagcat
     7561 actatggcaa aagcacatca ttcttccgtc aagaaagtca aaaacttctg caaagcgata
     7621 aaaaacgcac ggctgagtta gcaaacggcg ctctcggtat gattgagcta aacgatgatt
     7681 acacactgaa aaaagtgatg aaaccgctga ttgcatctaa cacagtaaca gatgaaattg
     7741 aacgcgcgaa cgtctttaaa atgaacggca aatggtacct gttcactgac tcccgcggat
     7801 caaaaatgac gattgacggc attacgtcta acgatattta catgcttggt tatgtttcta
     7861 attctttaac tggcccatac aagccgctga acaaaactgg ccttgtgtta aaaatggatc
     7921 ttgatcctaa cgatgtaacc tttacttact cacacttcgc tgtacctcaa gcgaaaggaa
     7981 acaatgtcgt gattacaagc tatatgacaa acagaggatt ctacgcagac aaacaatcaa
     8041 cgtttgcgcc aagcttcctg ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca
     8101 gcatccttga acaaggacaa ttaacagtta acaaataaaa acgcaaaaga aaatgccgat
     8161 ggccgcggta ccagatcttt aaatctagac ggtgaccaca acgcgcccgc tttgatcggg
     8221 gacgtctgcg gccgaccatt tacgggtctt gttgtcgttg gcggtcatgg gccgaacata
     8281 ctcacccgga tcggagggcc gaggacaagg tcgaacgagg ggcatgaccc ggtgcggggc
     8341 ttcttgcact cggcataggc gagtgctaag aataacgttg gcactcgcga ccggtgagtg
     8401 ctaggtcggg acggtgaggc caggcccgtc gtcgcagcga gtggcagcga ggacaacttg
     8461 agccgtccgt cgcgggcact gcgcccggcc agcgtaagta gcggggttgc cgtcacccgg
     8521 tgacccccgt ttcatccccg atccggagga atcacttcgc aatggccaag acaattgcgg
     8581 atccagctgc agaattt
//
