LOCUS       sequence-457353-        7550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is a BAC vector that contains both wt and mutant loxP sites.  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7550)
  AUTHORS   Coren JS
  TITLE     Retrofitting the BAC cloning vector pBeloBAC11 by the insertion of a
            mutant loxP site.
  JOURNAL   BMC Res Notes. 2017 Jul 28;10(1):344. doi: 
            10.1186/s13104-017-2631-8.
  PUBMED    28754135
REFERENCE   2  (bases 1 to 7550)
  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: "BMC Res 
            Notes."; date: "2017-07-28"; volume: "10(1):344. doi"; pages: " 
            10.1186/s13104-017-2631-8"
COMMENT     Sequence Label: pBeloBac11loxP2272
FEATURES             Location/Qualifiers
     source          1..7550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(56..436)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLL
                     THLCGISHRIWCTLSTICSDAA"
     primer_bind     61..80
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     274..296
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     288..305
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     289..305
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     312..331
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        312..330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    333..389
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(396..414)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(397..414)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(432..448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(432..448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(445..467)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    456..472
                     /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(480..486,487..504,505..510))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    525..546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    647..680
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(809..1468)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1383..1402
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1469..1571)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2498..2717
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             2808..3563
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    3566..3816
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             4142..5317
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             5317..6288
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    6361..6834
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    7094..7492
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(7510..7543)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gcggccgcaa ggggttcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
       61 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
      121 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
      181 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
      241 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
      301 ccagtgaatt gtaatacgac tcactatagg gcgaattcga gctcggtacc cggggatcct
      361 ctagagtcga cctgcaggca tgcaagcttg agtattctat agtgtcacct aaatagcttg
      421 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      481 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      541 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      601 cattaatgaa tcggccaacg cgaacccctt gcggccgccc ctcgagataa cttcgtataa
      661 agtatcctat acgaagttat cccgggccgt cgaccaattc tcatgtttga cagcttatca
      721 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcaa ccaggcgttt
      781 aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtact
      841 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
      901 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
      961 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     1021 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     1081 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     1141 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     1201 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg aattccggat
     1261 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     1321 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     1381 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     1441 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     1501 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     1561 cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag
     1621 ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcgcgat
     1681 aagctcatgg agcggcgtaa ccgtcgcaca ggaaggacag agaaagcgcg gatctgggaa
     1741 gtgacggaca gaacggtcag gacctggatt ggggaggcgg ttgccgccgc tgctgctgac
     1801 ggtgtgacgt tctctgttcc ggtcacacca catacgttcc gccattccta tgcgatgcac
     1861 atgctgtatg ccggtatacc gctgaaagtt ctgcaaagcc tgatgggaca taagtccatc
     1921 agttcaacgg aagtctacac gaaggttttt gcgctggatg tggctgcccg gcaccgggtg
     1981 cagtttgcga tgccggagtc tgatgcggtt gcgatgctga aacaattatc ctgagaataa
     2041 atgccttggc ctttatatgg aaatgtggaa ctgagtggat atgctgtttt tgtctgttaa
     2101 acagagaagc tggctgttat ccactgagaa gcgaacgaaa cagtcgggaa aatctcccat
     2161 tatcgtagag atccgcatta ttaatctcag gagcctgtgt agcgtttata ggaagtagtg
     2221 ttctgtcatg atgcctgcaa gcggtaacga aaacgatttg aatatgcctt caggaacaat
     2281 agaaatcttc gtgcggtgtt acgttgaagt ggagcggatt atgtcagcaa tggacagaac
     2341 aacctaatga acacagaacc atgatgtggt ctgtcctttt acagccagta gtgctcgccg
     2401 cagtcgagcg acagggcgaa gccctcgagt gagcgaggaa gcaccaggga acagcactta
     2461 tatattctgc ttacacacga tgcctgaaaa aacttccctt ggggttatcc acttatccac
     2521 ggggatattt ttataattat tttttttata gtttttagat cttctttttt agagcgcctt
     2581 gtaggccttt atccatgctg gttctagaga aggtgttgtg acaaattgcc ctttcagtgt
     2641 gacaaatcac cctcaaatga cagtcctgtc tgtgacaaat tgcccttaac cctgtgacaa
     2701 attgccctca gaagaagctg ttttttcaca aagttatccc tgcttattga ctctttttta
     2761 tttagtgtga caatctaaaa acttgtcaca cttcacatgg atctgtcatg gcggaaacag
     2821 cggttatcaa tcacaagaaa cgtaaaaata gcccgcgaat cgtccagtca aacgacctca
     2881 ctgaggcggc atatagtctc tcccgggatc aaaaacgtat gctgtatctg ttcgttgacc
     2941 agatcagaaa atctgatggc accctacagg aacatgacgg tatctgcgag atccatgttg
     3001 ctaaatatgc tgaaatattc ggattgacct ctgcggaagc cagtaaggat atacggcagg
     3061 cattgaagag tttcgcgggg aaggaagtgg ttttttatcg ccctgaagag gatgccggcg
     3121 atgaaaaagg ctatgaatct tttccttggt ttatcaaacg tgcgcacagt ccatccagag
     3181 ggctttacag tgtacatatc aacccatatc tcattccctt ctttatcggg ttacagaacc
     3241 ggtttacgca gtttcggctt agtgaaacaa aagaaatcac caatccgtat gccatgcgtt
     3301 tatacgaatc cctgtgtcag tatcgtaagc cggatggctc aggcatcgtc tctctgaaaa
     3361 tcgactggat catagagcgt taccagctgc ctcaaagtta ccagcgtatg cctgacttcc
     3421 gccgccgctt cctgcaggtc tgtgttaatg agatcaacag cagaactcca atgcgcctct
     3481 catacattga gaaaaagaaa ggccgccaga cgactcatat cgtattttcc ttccgcgata
     3541 tcacttccat gacgacagga tagtctgagg gttatctgtc acagatttga gggtggttcg
     3601 tcacatttgt tctgacctac tgagggtaat ttgtcacagt tttgctgttt ccttcagcct
     3661 gcatggattt tctcatactt tttgaactgt aatttttaag gaagccaaat ttgagggcag
     3721 tttgtcacag ttgatttcct tctctttccc ttcgtcatgt gacctgatat cgggggttag
     3781 ttcgtcatca ttgatgaggg ttgattatca cagtttatta ctctgaattg gctatccgcg
     3841 tgtgtacctc tacctggagt ttttcccacg gtggatattt cttcttgcgc tgagcgtaag
     3901 agctatctga cagaacagtt cttctttgct tcctcgccag ttcgctcgct atgctcggtt
     3961 acacggctgc ggcgagcgct agtgataata agtgactgag gtatgtgctc ttcttatctc
     4021 cttttgtagt gttgctctta ttttaaacaa ctttgcggtt ttttgatgac tttgcgattt
     4081 tgttgttgct ttgcagtaaa ttgcaagatt taataaaaaa acgcaaagca atgattaaag
     4141 gatgttcaga atgaaactca tggaaacact taaccagtgc ataaacgctg gtcatgaaat
     4201 gacgaaggct atcgccattg cacagtttaa tgatgacagc ccggaagcga ggaaaataac
     4261 ccggcgctgg agaataggtg aagcagcgga tttagttggg gtttcttctc aggctatcag
     4321 agatgccgag aaagcagggc gactaccgca cccggatatg gaaattcgag gacgggttga
     4381 gcaacgtgtt ggttatacaa ttgaacaaat taatcatatg cgtgatgtgt ttggtacgcg
     4441 attgcgacgt gctgaagacg tatttccacc ggtgatcggg gttgctgccc ataaaggtgg
     4501 cgtttacaaa acctcagttt ctgttcatct tgctcaggat ctggctctga aggggctacg
     4561 tgttttgctc gtggaaggta acgaccccca gggaacagcc tcaatgtatc acggatgggt
     4621 accagatctt catattcatg cagaagacac tctcctgcct ttctatcttg gggaaaagga
     4681 cgatgtcact tatgcaataa agcccacttg ctggccgggg cttgacatta ttccttcctg
     4741 tctggctctg caccgtattg aaactgagtt aatgggcaaa tttgatgaag gtaaactgcc
     4801 caccgatcca cacctgatgc tccgactggc cattgaaact gttgctcatg actatgatgt
     4861 catagttatt gacagcgcgc ctaacctggg tatcggcacg attaatgtcg tatgtgctgc
     4921 tgatgtgctg attgttccca cgcctgctga gttgtttgac tacacctccg cactgcagtt
     4981 tttcgatatg cttcgtgatc tgctcaagaa cgttgatctt aaagggttcg agcctgatgt
     5041 acgtattttg cttaccaaat acagcaatag taatggctct cagtccccgt ggatggagga
     5101 gcaaattcgg gatgcctggg gaagcatggt tctaaaaaat gttgtacgtg aaacggatga
     5161 agttggtaaa ggtcagatcc ggatgagaac tgtttttgaa caggccattg atcaacgctc
     5221 ttcaactggt gcctggagaa atgctctttc tatttgggaa cctgtctgca atgaaatttt
     5281 cgatcgtctg attaaaccac gctgggagat tagataatga agcgtgcgcc tgttattcca
     5341 aaacatacgc tcaatactca accggttgaa gatacttcgt tatcgacacc agctgccccg
     5401 atggtggatt cgttaattgc gcgcgtagga gtaatggctc gcggtaatgc cattactttg
     5461 cctgtatgtg gtcgggatgt gaagtttact cttgaagtgc tccggggtga tagtgttgag
     5521 aagacctctc gggtatggtc aggtaatgaa cgtgaccagg agctgcttac tgaggacgca
     5581 ctggatgatc tcatcccttc ttttctactg actggtcaac agacaccggc gttcggtcga
     5641 agagtatctg gtgtcataga aattgccgat gggagtcgcc gtcgtaaagc tgctgcactt
     5701 accgaaagtg attatcgtgt tctggttggc gagctggatg atgagcagat ggctgcatta
     5761 tccagattgg gtaacgatta tcgcccaaca agtgcttatg aacgtggtca gcgttatgca
     5821 agccgattgc agaatgaatt tgctggaaat atttctgcgc tggctgatgc ggaaaatatt
     5881 tcacgtaaga ttattacccg ctgtatcaac accgccaaat tgcctaaatc agttgttgct
     5941 cttttttctc accccggtga actatctgcc cggtcaggtg atgcacttca aaaagccttt
     6001 acagataaag aggaattact taagcagcag gcatctaacc ttcatgagca gaaaaaagct
     6061 ggggtgatat ttgaagctga agaagttatc actcttttaa cttctgtgct taaaacgtca
     6121 tctgcatcaa gaactagttt aagctcacga catcagtttg ctcctggagc gacagtattg
     6181 tataagggcg ataaaatggt gcttaacctg gacaggtctc gtgttccaac tgagtgtata
     6241 gagaaaattg aggccattct taaggaactt gaaaagccag caccctgatg cgaccacgtt
     6301 ttagtctacg tttatctgtc tttacttaat gtcctttgtt acaggccaga aagcataact
     6361 ggcctgaata ttctctctgg gcccactgtt ccacttgtat cgtcggtctg ataatcagac
     6421 tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc acggtcccac
     6481 tcgtatcgtc ggtctgatta ttagtctggg accacggtcc cactcgtatc gtcggtctga
     6541 taatcagact gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca
     6601 tggtcccact cgtatcgtcg gtctgattat tagtctggga ccacggtccc actcgtatcg
     6661 tcggtctgat tattagtctg gaaccacggt cccactcgta tcgtcggtct gattattagt
     6721 ctgggaccac ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacgatccca
     6781 ctcgtgttgt cggtctgatt atcggtctgg gaccacggtc ccacttgtat tgtcgatcag
     6841 actatcagcg tgagactacg attccatcaa tgcctgtcaa gggcaagtat tgacatgtcg
     6901 tcgtaacctg tagaacggag taacctcggt gtgcggttgt atgcctgctg tggattgctg
     6961 ctgtgtcctg cttatccaca acattttgcg cacggttatg tggacaaaat acctggttac
     7021 ccaggccgtg ccggcacgtt aaccgggctg catccgatgc aagtgtgtcg ctgtcgacga
     7081 gctcgcgagc tcggacatga ggttgccccg tattcagtgt cgctgatttg tattgtctga
     7141 agttgttttt acgttaagtt gatgcagatc aattaatacg atacctgcgt cataattgat
     7201 tatttgacgt ggtttgatgg cctccacgca cgttgtgata tgtagatgat aatcattatc
     7261 actttacggg tcctttccgg tgatccgaca ggttacgggg cggcgacctc gcgggttttc
     7321 gctatttatg aaaattttcc ggtttaaggc gtttccgttc ttcttcgtca taacttaatg
     7381 tttttattta aaataccctc tgaaaagaaa ggaaacgaca ggtgctgaaa gcgagctttt
     7441 tggcctctgt cgtttccttt ctctgttttt gtccgtggaa tgaacaatgg aagtccgagc
     7501 tcatcgctaa taacttcgta tagcatacat tatacgaagt tatattcgat
//
