LOCUS       40924_45258        3695 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gene deletion vector pUCGM-lox, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3695)
  AUTHORS   Quenee L, Lamotte D, Polack B.
  TITLE     Combined sacB-based negative selection and cre-lox antibiotic marker
            recycling for efficient gene deletion in pseudomonas aeruginosa
  JOURNAL   BioTechniques 38 (1), 63-67 (2005)
  PUBMED    15679087
REFERENCE   2  (bases 1 to 3695)
  AUTHORS   Quenee L, Lamotte D, Polack B.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JAN-2005) GREPI, CHU, Hopital Michallon, Grenoble 
            38043, France
REFERENCE   3  (bases 1 to 3695)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3695)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "2005"; volume: "38"; issue: "1"; pages: 
            "63-67"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-JAN-2005) GREPI, CHU, Hopital Michallon, Grenoble 38043, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    492..525
                     /label=Cre recombinase binding site
                     /bound_moiety="Cre recombinase"
                     /note="loxP, Cre recombinase recognition site"
     promoter        534..562
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             751..1281
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(1328..1361)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1474..1490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1498..1514)
                     /label=lac operator
                     /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(1522..1552)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1567..1588)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2638..3495)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3496..3600)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atccccgggt accgagctct tcagctgtac aattggtacc atggatgcat
      481 atggcggccg cataacttcg tatagcatac attatacgaa gttatctagc gcgtcgacat
      541 aagcctgttc ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
      601 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
      661 actgtttttt tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
      721 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagcaacga tgttacgcag
      781 cagggcagtc gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt
      841 aggctcggcc ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag
      901 ttcggagacg tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg
      961 ctccgtagta agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct
     1021 ctcgcggctt acgttctgcc caggtttgag cagccgcgta gtgagatcta tatctatgat
     1081 ctcgcagtct ccggcgagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc
     1141 aagcatgagg ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac
     1201 gatcccgcag tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat
     1261 atcgacccaa gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgac
     1321 gcgctagata acttcgtata gcatacatta tacgaagtta tggatccagc ttatcgatac
     1381 cgcgggccct acgtacgcgt gttaaccggt gagctcggta cccggggatc ctctagagtc
     1441 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1501 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1561 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1621 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2281 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     2341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2761 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3601 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3661 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
