LOCUS       sequence-146415-        5120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Mu-gam under the pBAD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5120)
  AUTHORS   Cui L, Bikard D
  TITLE     Consequences of Cas9 cleavage in the chromosome of Escherichia coli.
  JOURNAL   Nucleic Acids Res. 2016 Apr 8. pii: gkw223.
  PUBMED    27060147
REFERENCE   2  (bases 1 to 5120)
  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: "Nucleic 
            Acids Res. 2016 Apr 8. pii: gkw223."
COMMENT     Sequence Label: pLC13
FEATURES             Location/Qualifiers
     source          1..5120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(1902..1919)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2072..2158
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2250..2277
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2296..2387
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2388..2456,2457..3248)
                     /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(2606..2625)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3290..3745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3377..3396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3856..4444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4345..4364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4598..4615
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4630..4770
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4696..4715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4856..4878
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5017..5036
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcgaatt cgagctcggt
     1321 aactttaaga aggagatata ccatggctaa accagcaaaa cgtatcaaga gtgccgcagc
     1381 ggcttatgtg ccacaaaacc gcgatgcggt gattaccgat attaaacgca tcggggattt
     1441 acagcgcgaa gcatcacgtc tggaaacgga aatgaatgat gccatcgcgg aaattacgga
     1501 gaaatttgcg gcccggattg caccgattaa aaccgatatt gaaacccttt caaaaggcgt
     1561 tcagggatgg tgtgaagcga accgcgacga actgacgaac ggcggcaaag tgaagacggc
     1621 gaatcttgtc accggtgatg tatcgtggcg ggtccgtcca ccatcagtaa gtattcgtgg
     1681 tatggatgca gtgatggaaa cgctggagcg tcttggcctg caacgcttta ttcgcacgaa
     1741 gcaggaaatc aacaaggaag cgattttact ggaaccgaaa gcggtcgcag gcgttgccgg
     1801 aattacagtt aaatcaggca ttgaggattt ttctattatt ccatttgaac aggaagccgg
     1861 tatttaattg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     1921 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     1981 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     2041 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2101 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2161 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     2221 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     2281 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2341 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2401 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2461 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2521 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2581 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     2641 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2701 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2761 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2821 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2881 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     2941 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3001 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3061 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3121 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3181 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3241 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt
     3301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3361 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3481 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     3541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3601 caaacttgaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     3661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3721 aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct
     3781 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3841 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3901 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3961 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     4021 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4081 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4141 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4201 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4261 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4321 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4381 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4441 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4501 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4561 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4621 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4681 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4741 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4801 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4861 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcaaggagat
     4921 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     4981 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     5041 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
     5101 ctcatgtttg acagcttatc
//
