LOCUS       sequence-344536-        8380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAPTURE method receiver plasmid for Bacillus cloning and 
            heterologous expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8380)
  AUTHORS   Enghiad B, Huang C, Guo F, Jiang G, Wang B, Tabatabaei SK, Martin 
            TA, Zhao H
  TITLE     Cas12a-assisted precise targeted cloning using in vivo Cre-lox 
            recombination.
  JOURNAL   Nat Commun. 2021 Feb 19;12(1):1171. doi: 10.1038/s41467-021-21275-4.
  PUBMED    33608525
REFERENCE   2  (bases 1 to 8380)
  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: "Nat 
            Commun."; date: "2021-02-19"; volume: "12(1):1171. doi"; pages: " 
            10.1038/s41467-021-21275-4"
COMMENT     Sequence Label: pBE50
FEATURES             Location/Qualifiers
     source          1..8380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1608..1946)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSISRAAADPLESTCSPSLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRINSPMSSTSGIG
                     SAADAKCR"
     primer_bind     1847..1869
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1861..1878
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1862..1878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1947..1963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1947..1963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1960..1982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1971..1987
                     /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)."
     promoter        complement(join(1995..2001,2002..2019,2020..2025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2040..2061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2774..2792)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2860..2964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2965..3033,3034..3825)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3183..3202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    complement(3963..3996)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(5451..5484)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      5585..6130
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     6031..6050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6247..6264
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggataaagca cagctacaga cctggcattg atcgtgcctg tcagtttacc atcgttcact
       61 tgaaatgaac ccgctccagc tttattgtca tacctgccat caggcaattt tgttgccgta
      121 ttgatagaga cagaggatga acctgcattt gccagcacaa cgccatgtga gccgcgctga
      181 ttcataaata tctggttgtt tccattcggg ttcgagagtt cctcaggctg tccagccatc
      241 acattgtgaa atctattgac cgcagtgata gcctgatctt caaataaagc actcccgcga
      301 tcgcctattt ggcttttccc cgggaacctc acaccatttc cgcctccctc aggtctggaa
      361 aagaaaagag gcgtactgcc tgaacgagaa gctatcaccg cccagcctaa acggattccc
      421 cctatgcaag ggtttattgt tttctaaaat ctgattacca attagaatga atatttccca
      481 aatattaaat aataaaacaa aaaaattgaa aaaagtgttt ccaccatttt ttcaattttt
      541 ttataatttt tttaatctgt tatttaaata gtttatagtt aaatttacat tttcattagt
      601 ccattcaata ttctctccaa gataactacg aactgctaac aaaattctct ccctatgttc
      661 taatggagaa gattcagcca ctgcatttcc cgcaatatct tttggtatga ttttacccgt
      721 gtccatagtt aaaatcatac ggcataaagt taatatagag ttggtttcat catcctgata
      781 attatctatt aattcctctg acgaatccat aatggctctt ctcacatcag aaaatggaat
      841 atcaggtagt aattcctcta agtcataatt tccgtatatt cttttatttt ttcgttttgc
      901 ttggtaaagc attatggtta aatctgaatt taattccttc tgaggaatgt atccttgttc
      961 ataaagctct tgtaaccatt ctccataaat aaattcttgt ttgggaggat gattccacgg
     1021 taccatttct tgctgaataa taattgttaa ttcaatatat cgtaagttgc ttttatctcc
     1081 tatttttttt gaaataggtc taattttttg tataagtatt tctttacttt gatctgtcaa
     1141 tggttcagat acgacgacta aaaagtcaag atcactattt ggttttagtc cactctcaac
     1201 tcctgatcca aacatgtaag taccaataag gttatttttt aaatgtttcc gaagtatttt
     1261 tttcacttta ttaatttgtt cgtatgtatt caaatatatc ctcctcacta ttttgattag
     1321 tacctatttt atatccatag ttgttaatta aataaactta atttagttta tttatagatt
     1381 tcattggctt ctaaattttt tatctagata ataattattt tagttaattt tattctagat
     1441 tatatatgat atgatctttc atttccataa aactaaagta agtgtaaacc tattcattgt
     1501 tttaaaaata tctcttgcca gtcacgttac gttattagtg taggagggcg tggatatgtc
     1561 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     1621 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaatttat gcggtgtgaa
     1681 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     1741 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     1801 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     1861 tgtaaaacga cggccagtgc caagcttggg ctgcaggtcg actctagagg atccgcggcc
     1921 gcgcgcgata tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     1981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgctacca
     2101 ttaccagttg gtctggtgtc aaaaataata ataaccgggc aggccatgtc tgcccgtatt
     2161 tcgcgtaagg aaatccatta tgtactattt aattctgcgt gacatcccat cgatcagacc
     2221 agtttttaat ttgtgtgttt ccatgtgtcc agtttggaat actcttaacc tcattggaaa
     2281 tcgcggcata atcactggtg gtatgattga tgaccgcgtc aacaatgacc tttatgccat
     2341 attcttcagc ggctgcacac atttctttaa attcttgttc agtacctaag taacggttgc
     2401 caatttgata cgatgtcggc tgatacagcc agtaccagtt cgacatgctt ttatctcctt
     2461 gattcccttc ctttacttgg ttaatcggag atgtctgaat ggctgtatat cctgcatcat
     2521 gaatatcctt catattgtgt tttaacgtat tgaacgacca attccatgca tgaagaatgg
     2581 ttccgctttt gatcgacggt gctgtaagct cattcgattt gttcgccgtt tcagcactcg
     2641 cagccgccgg tcctgccaga accaaatgaa acagcaataa aaatccagcg aataacggca
     2701 gtaaagaggt tttgaatcgt tttgcaaaca ttcttgacac tccttatttg attttttgaa
     2761 gaaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2821 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2881 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2941 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3001 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3061 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3121 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3181 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     3241 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3301 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3361 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3421 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3481 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     3541 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3601 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3661 tctggagccg gtgagcgtgg gtcacgcggt atcattgcag cactggggcc agatggtaag
     3721 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3781 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3841 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3901 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3961 gcataacttc gtatagcata cattatacga agttatgatc ctccagcatc aaacctgctg
     4021 tcgcttttgc cgagccgatg acgccaggaa tgcctgcgcc gggatgcgtg cctgcgccga
     4081 ccaggtagag attagtaatg gttttatcgc ggttatgcgg ccgaaaccag gcgctctggg
     4141 taagaacggg ctccacagaa aaggctgagc catgataggc attaagctgg tcgcgaaaat
     4201 caaacggcgt aaacatccgg tgcgtgacca gctgactccg taagccaggc atgtaatgct
     4261 gctcaaggta cgcaaaaata cggtcgcgta gttttggccc ctcaaccgtc cagtcgaggt
     4321 tcgcggtgcc taaatgcggc accggcgcca acacatagta actgccgcaa ccttcaggcg
     4381 ccagtgacga atccgtgaca cagggcgcgt gcagataaag tgagaagtcc tctgcgaggc
     4441 catcatgatt aaaaatttcg tcaatcagct cgcggtaacg cgggccgaaa caaaccgtgt
     4501 gatgcgcgag ctgatcatga tggtgattca aaccaaaata gagcacaaac agagagttac
     4561 tcatgcgctt agtctgcagt ttgttggact gcttaaccgc ggcagggtgc tggcttaaca
     4621 ggtcgcgata ggtatgaacc acatctgcat ttgacgcgac ggcttgcgtc aggaacctgc
     4681 gaccgtcctc taaatgcacg gcttcaatct tgtttcctgt cgtttccata tggctgactc
     4741 tggcgtttaa cacgacttcg ccacccagat cctgaaacag ctttatcatc ccctgaacta
     4801 atgcgccggt gccgccacgc ggaaaccaga cgccccactc acgctccagc gcgtgtatca
     4861 acgtataaat ggatgaggtg gcgaagggat tgccgcccac caacagcgag tggaaagaaa
     4921 acgcctggcg cagatgttca tcttcgatgt aactggcaac cttactgtaa acgcttctcc
     4981 atgcctgcag tttcgccagt tgaggtgcgg cgcgaagcat gtctctgaac gataaaaaag
     5041 ggacagtacc gagctttaga tagccttctt taaacaccgc gcgtgaatag tccagaaact
     5101 gacgataacc ttcgacatcg cggggattaa actgctgaat ctgcgcttcg agccgggttt
     5161 gatcgttatc gtaattaaag accttccctg actcccaaca caggcggtaa aacggcgtaa
     5221 ccggcagcag ttcgacatac tcttttaact gttttcctgc cagtgcaaac agttcttcaa
     5281 tggcactggg atcggtgata accgtcgggc ctgcatcaaa ggtaaacccc tgatcctcgt
     5341 agacataagc ccgaccgccg ggtttatcac gttgttcaag cagtaagacg gggatccccg
     5401 cagcttgtag acgaattgcc agtgccaggc caccgaagcc tgcaccaatt ataacttcgt
     5461 atagcataca ttatacgaag ttatacaact tatatcgtat ggggctgact tcaggtgcta
     5521 catttgaaga gataaattgc actgaaatct agaaatattt tatctgatta ataagatgat
     5581 cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct
     5641 tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc
     5701 ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact
     5761 tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt
     5821 ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg
     5881 gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg
     5941 cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg
     6001 aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt
     6061 cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag
     6121 cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca
     6181 tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc
     6241 gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga
     6301 cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag
     6361 tgccaacata gtaagccagt atacactccg ctagcgctga tgtccggcgg tgcttttgcc
     6421 gttacgcacc accccgtcag tagctgaaac cgcctctccc cgcgcgttgg ccgattcatt
     6481 aatgcagctg gcacgacagg tttcccgacg acctcgagct ggatacttcc cgtccgccag
     6541 ggggacatgc cggcgatgct gaaggtcgcg cgcattcccg atgaagaggc cggttaccgc
     6601 ctgtttgagg atatagtaat ctttctaaat agctttggat tggaggagta tggccactaa
     6661 tactaagttc agctaataaa aaaatttgct aaagaactcc agctggattt cactgatgag
     6721 aatatcgtcg gagataaata taataattcc acggactata gactatacta gtatactccg
     6781 tctactgtac gatacacttc cgctcaggtc cttgtccttt aacgaggatt gttacgtacg
     6841 ctaatggcgt caaaacaaag actctagacc taggccttaa gatctgatca tatgcatccg
     6901 cgggcccggg ttaacgcgta atccatggat caagagacag gatgaggatc gtttcgcatg
     6961 attgaacaag atggattgca cgcaggttct ccggtgccct gaatgaactg cagaaagagc
     7021 tggtagttgg cgcactgttc gaagaactgc cgatgtccag taagattctt actatgctgg
     7081 ttgaaccgga tgctggtaaa gctacttggg ttgctgcttc tacttatggt accgatacaa
     7141 ctactggtga ggaagttaaa ggagctctta aagaaatcca cttcagtagg tggcccggct
     7201 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     7261 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     7321 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     7381 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     7441 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgactaaga aaatgccgtc
     7501 aaatccgctc gccatgactt cactaacgat gcctttgaaa atcttcaagt tcttttctac
     7561 taattcaagg cgtgtctcac caggtttttg gtttgctccg gcgcaaatgc agacaatatc
     7621 agcatccttg cagggtatgt ttctctttga tgtctttttg tttgtgaagt atttcacatt
     7681 tatattgtgc aacacttcac aaacttttgc aagagaaaag ttttgtctga tttatgaaca
     7741 aaaaagaaac catcattgat ggtttctttc ggtaagtccc gtctagcctt gccctcaatg
     7801 gggaagagaa ccgcttaagc ccgagtcatt atataaacca tttagcacgt aatcaaagcc
     7861 aggctgattc tgaccgggca cttgggcgct gccattatta aaaatcactt ttgcgttggt
     7921 tgtatccgtg tccgcaggca gcgtcagcgt gtaaattccg tctgcatttt tagtcattgg
     7981 ttttccaggc caagatccgg tcaattcaat tactcggctc ccatcatgtt tatagatata
     8041 agcatttacc tggctccaat gattcggatt ttgatagccg atggttttgg ccgacgctgg
     8101 atctctttta acaaaactgt atttctcggt cctcgttaca ccatcactgt tcgttccttt
     8161 taacatgatg gtgtatgttt tgccaaatgg atctcctttt ccgattgtga attgatctcc
     8221 atccttaaac gccgtctctg gtccattatt gatttgataa acggcttttg ttgtattcgc
     8281 atctgcacgc aaggtaatcg tcagttgatc attgaaagaa tgtgttacac ctgttttgta
     8341 attctcaagg aaaacatgag gcgcttttgc aatatcatca
//
