LOCUS       sequence-11290-e        6594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6594)
  TITLE     George Cross Lab Plasmids
REFERENCE   2  (bases 1 to 6594)
  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"
COMMENT     Sequence Label: pLEW100cre_del_tetO
FEATURES             Location/Qualifiers
     source          1..6594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1082..1456)
                     /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"
     primer_bind     complement(1595..1614)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1596..1614)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2054..3085
                     /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"
     promoter        complement(3830..3848)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3831..3848)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4139..4158)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4369..4388)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4488..4510
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4548..4566)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4634..4738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4739..4807,4808..5599)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHSMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4957..4976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5770..6358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6259..6278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6512..6529
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaattcgagc tcatatagtt ggtatgtatt ctaattccag actactggcg tggataaaca
       61 tgtcccctga ttaaaaggaa agattccata gcccattagt gcaaagataa ttggtacacc
      121 gtcaaaaaca catgggagac cacacgatac gacactgtga ccgtagcatc aattccgcac
      181 tcgatataac tctatcgaag aacttccatg gtacaaactg gtcgccacgg tctgctccac
      241 acggagatca tcgtatcatt tttatcgata gcggccgcta tcgatgtatg ccttggccct
      301 gatggcatgc caatttcact acaacggact atgtggaccc cgttatcatg gaaatgcgct
      361 agttggagga agttagaccg cgccggaaaa gagaggggta gagaaaatga caacttggaa
      421 gatatccaca cacgcacggt gaaacgttag caacaattat tagggaagca cgcttgccct
      481 agtcccacga gtaaccaaga ctccaaaagc ctttctggca cagagagcga gccgaaatgg
      541 aaaagagaaa caatgcctgc actaacacta ctgagcgatt cgcctcgccg cggaggaccg
      601 aatactaata acgacacttg cggtcaaaaa gtagaagaac aaatgctcaa cgatgagtga
      661 atcaggttag ggtagttgga aaattataca gaatgtcttt ggcaacacac cggcnnnctt
      721 gcatgcctgc aaggccttgc agaatactgc atagataaca aacgcatcaa acacaactgg
      781 tggcactatt tcaatcatgt cgacacacca agacgagaaa atttgagtta aacaagatgc
      841 ctacaaccct atatttatta tcttcaagtg atcttctctt ctcttgtgcg ctgtacgtaa
      901 atgtgttgca aataaggcat agggctgagt acaggcacca cacgcgagga aaatcataag
      961 cactagccgt gtcaactcac agcgactaca gaacaatttt ggccacacaa cccggtgtta
     1021 ggatctccga ggcctgggac ccgtgggccg ccgtcggacc ggcggtgttg gtcggcgtcg
     1081 gtcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga
     1141 actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga
     1201 agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca
     1261 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca
     1321 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc
     1381 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac
     1441 tggtcaactt ggccatggtg atagcttatt ttatggcagc aacgagacct tacgtaaagc
     1501 aaaactaaat aagagcggag actgcaatgc agagtaaaaa aaaaaaaaac atgaatttca
     1561 gaagaccttg ctgtgccccc ggtacgggag atctccctat agtgagtcgt attaatcagg
     1621 taccgtcatt ggggttaagc ggaaaggtgt gtgtcagtag gttgtgaggt gaaagcgttt
     1681 tcagatgcat agtgagctta atgtcctttt cacagtatat cgtgtctgat aggtatctct
     1741 tattagtata gtcgaatact agtcaatagt gcgttttgtg caaaatgtcc attttgtggc
     1801 agtgatgggg ttgttttatg ctattccgtg tctctgggtg ggcgtgcatt gaaaataggg
     1861 gttatcgggt gagggatcga gtactgagtt taacatgttc tcgtcccggg ctgcacgcgc
     1921 cttcgagttt tttttccttt tccccatttt tttcaacttg aagacttcaa ttacaccaaa
     1981 aagtaaaatt cacaagctcg aggggcagag ccgatcctgt acactttact taaaaccatt
     2041 atctgagtgt gaaatgtcca atttactgac cgtacaccaa aatttgcctg cattaccggt
     2101 cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg atcgccaggc
     2161 gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg
     2221 caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg attatcttct
     2281 atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg gccagctaaa
     2341 catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg tttcactggt
     2401 tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc
     2461 gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg atcgctgcca
     2521 ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac gtatagccga
     2581 aattgccagg atcagggtta aagatatctc acgtactgac ggtgggagaa tgttaatcca
     2641 tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac ttagcctggg
     2701 ggtaactaaa ctggtcgagc gatggatttc cgtctctggt gtagctgatg atccgaataa
     2761 ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct
     2821 atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt acggcgctaa
     2881 ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg tcggagccgc
     2941 gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg gtggctggac
     3001 caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag gggcaatggt
     3061 gcgcctgctg gaagatggcg attagccatt aacgcggatc ctgcccattt agttggcttt
     3121 tcccttgtct cgtgtctttt ccgtggaaag gttcccggag taatctgatg gcacagcagg
     3181 gaggtgcgcc tgcaggttgg ttaggaaggg gggatgatgt aaaagaagaa aatgggggga
     3241 tatcttatgt ttaagaggat aaataatgtg aaggggcttt atatgcttgc ttggttgtct
     3301 cgttggtgca tggggaatct gcatgtttgc tttggagcac gcgtggtaca tcaatgggat
     3361 catatcttgc tgcctcccgc agctcaccgt gcgagctgcc gggaccccgt ttctcatagg
     3421 cgtgtgcacc ttccgtctga gacctgtaaa tggttaaaaa ggaatatata attacctttt
     3481 gaaaagtggt aaaaaacgaa tatatctttt tttttggttt taatacgctc ttttgtgtgt
     3541 atgagaggaa taaataagtg tgtgtgtgtg tgtgtgttgt tgttgttgaa aaaatgaagg
     3601 acacgaaagc gctcattagg ggtggggata caccaggcat aattatccga aaatattttg
     3661 atgcaccaaa taagtgaaca tactgacgaa atcaaagagt tggaggatgg atagggaggc
     3721 ctcattggca gtggtaaatt gatttgactt gaatatgata ccctcttact gtttcattgc
     3781 cactacaagt tggtttcctt cccctgcagg catgcaagct agcttgtatt ctatagtgtc
     3841 acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg
     3901 acaatatgta caagcctaat tgtgtagcat ctggcttact gaagcagacc ctatcatctc
     3961 tctcgtaaac tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt tctggtaacg
     4021 ccgttccgca ccccggaaat ggtcagcgaa ccaatcagca gggtcatcgc tagccagatc
     4081 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     4141 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     4201 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     4261 ttgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
     4321 taatgcagga gtcgcataag ggagagcgtc gatatggtgc actctcagta caatctgctc
     4381 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4441 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4501 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4561 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4621 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4741 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4801 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4861 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4921 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4981 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5041 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5101 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5161 aggaccgaag gagctaaccg cttttttgca cagcatgggg gatcatgtaa ctcgccttga
     5221 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5281 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5341 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5401 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5461 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5521 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5581 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5641 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5701 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5761 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5821 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5881 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6241 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6301 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6361 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6421 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6481 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6541 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcag
//
