LOCUS       sequence-39752-e        7685 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 7685)
  TITLE     George Cross lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7685)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLEW100v5-trunc
FEATURES             Location/Qualifiers
     source          1..7685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(757..804)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      complement(889..936)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1345..1719)
                     /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(1857..1876)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1858..1876)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2153..2171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2174..2192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2388..4040
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKRLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAK*GGKSKL"
     primer_bind     complement(2454..2472)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(4924..4942)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4925..4942)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5232..5251)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5461..5480)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5640..5658)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5726..5830
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5831..5899,5900..6691)
                     /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(6049..6068)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6862..7450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7351..7370
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7604..7621
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaattcgagc tccagctgag ctcatatagt tggtatgtat tctaattcca gactactggc
       61 gtggataaac atgtcccctg attaaaagga aagattccat agcccattag tgcaaagata
      121 attggtacac cgtcaaaaac acatgggaga ccacacgata cgacactgtg accgtagcat
      181 caattccgca ctcgatataa ctctatcgaa gaacttccat ggtacaaact ggtcgccacg
      241 gtctgctcca cacggagatc atcgtatcat ttttatcgat agcggccgct atcgatgtat
      301 gccttggccc tgatggcatg ccaatttcac tacaacggac tatgtggacc ccgttatcat
      361 ggaaatgcgc tagttggagg aagttagacc gcgccggaaa agagaggggt agagaaaatg
      421 acaacttgga agatatccac acacgcacgg tgaaacgtta gcaacaatta ttagggaagc
      481 acgcttgccc tagtcccacg agtaaccaag actccaaaag cctttctggc acagagagcg
      541 agccgaaatg gaaaagagaa acaatgcctg cactaacact actgagcgat tcgcctcgcc
      601 gcggaggacc gaatactaat aacgacactt gcggtcaaaa agtagaagaa caaatgctca
      661 acgatgagtg aatcaggtta gggtagttgg aaaattatac agaatgtctt tggcaacaca
      721 ccgcttaagc ctatccggat atagttctcc tttcagcaaa aaacccctca agacccgttt
      781 agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc
      841 ctttcgggct ttgttagcag ccctatccgg atatagttct cctttcagca aaaaacccct
      901 caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc
      961 caactcagct tcctttcggg ctttgttagc agcatttaaa tgcagaatac tgcatagata
     1021 acaaacgcat caaacacaac tggtggcact atttcaatca tgtcgacaca ccaagacgag
     1081 aaaatttgag ttaaacaaga tgcctacaac cctatattta ttatcttcaa gtgatcttct
     1141 cttctcttgt gcgctgtacg taaatgtgtt gcaaataagg catagggctg agtacaggca
     1201 ccacacgcgt ggaaaatcat aagcactagc cgtgtcaact cacagcgact acagaacaat
     1261 tttggccaca caacccggtg ttaggatctc cgaggcctcg gacccgtcgg gccgcgtcgg
     1321 accggcggtg ttggtcggcg tcggtcagtc ctgctcctcg gccacgaagt gcacgcagtt
     1381 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     1441 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     1501 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     1561 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     1621 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     1681 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtgatagctt attttatggc
     1741 agcaacgaga ccttacgtaa agcaaaacta aataagagcg gagactgcaa tgcagagtaa
     1801 aaaaaaaaaa acatgaattt cagaagacct tgctgtgccc ccggtacggg agatctccct
     1861 atagtgagtc gtattaatca ggtacctagc tttccaccca gcgcgggtgc attctggctc
     1921 ttatatatac ttattgtcat gacagagtat attgtactgt gttgataagg gacgggtaac
     1981 tgtattgaag agccgatgct tttgacatgt tagatataat atgttttatt gtaaagtcaa
     2041 tacaacacac aataggataa taatgataaa gttaaaaaag tatatatagt aatagaaata
     2101 tatcttatat aggaaagatt aagcagtaaa agtagcgctt acggcgtacg gttccctatc
     2161 agtgatagag atctccctat cagtgataga gatccctgag tactgagttt aacatgttct
     2221 cgtcccgggc tgcacgcgcc ttcgagtttt ttttcctttt ccccattttt ttcaacttga
     2281 agacttcaat tacaccaaaa agtaaaattc acaagcttgg aattcctttg tgttacattc
     2341 ttgaatgtcg ctcgcagtga cattagcatt ccggtactgt tggtaaaatg gaagacgcca
     2401 aaaacataaa gaaaggcccg gcgccattct atcctctaga ggatggaacc gctggagagc
     2461 aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg
     2521 cacatatcga ggtgaacatc acgtacgcgg aatacttcga aatgtccgtt cggttggcag
     2581 aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact
     2641 ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg
     2701 cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gaacatttcg cagcctaccg
     2761 tagtgtttgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aaattaccaa
     2821 taatccagaa aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt
     2881 acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt gtaccagagt
     2941 cctttgatcg tgacaaaaca attgcactga taatgaattc ctctggatct actgggttac
     3001 ctaagggtgt ggcccttccg catagaactg cctgcgtcag attctcgcat gccagagatc
     3061 ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc
     3121 acggttttgg aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa
     3181 tgtatagatt tgaagaagag ctgtttttac gatcccttca ggattacaaa attcaaagtg
     3241 cgttgctagt accaacccta ttttcattct tcgccaaaag cactctgatt gacaaatacg
     3301 atttatctaa tttacacgaa attgcttctg ggggcgcacc tctttcgaaa gaagtcgggg
     3361 aagcggttgc aaaacgcttc catcttccag ggatacgaca aggatatggg ctcactgaga
     3421 ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag
     3481 ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta
     3541 atcagagagg cgaattatgt gtcagaggac ctatgattat gtccggttat gtaaacaatc
     3601 cggaagcgac caacgccttg attgacaagg atggatggct acattctgga gacatagctt
     3661 actgggacga agacgaacac ttcttcatag ttgaccgctt gaagtcttta attaaataca
     3721 aaggatatca ggtggccccc gctgaattgg aatcgatatt gttacaacac cccaacatct
     3781 tcgacgcggg cgtggcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg
     3841 ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc
     3901 aagtaacaac cgcgaaaagg ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag
     3961 gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagtagg
     4021 gcggaaagtc caaattgtaa aatgtaactg tattcagcga tgacgaaatt cttagctatt
     4081 gtaatattat atgcaaattg atgaatggta attttgtaat tgtgggtcac tgtactattt
     4141 taacgaataa taaaatcagg tataggtaac taaaaaggaa ttcgagctcg gatcctgccc
     4201 atttagttgg cttttccctt gtctcgtgtc ttttccgtgg aaaggttccc ggagtaatct
     4261 gatggcacag cagggaggtg cgcctgcagg ttggttagga aggggggatg atgtaaaaga
     4321 agaaaatggg gggatatctt atgtttaaga ggataaataa tgtgaagggg ctttatatgc
     4381 ttgcttggtt gtctcgttgg tgcatgggga atctgcatgt ttgctttgga gcacgcgtgg
     4441 tacatcaatg ggatcatatc ttgctgcctc ccgcagtcac cgtgcggagc tgccggtgcc
     4501 ccgtttctca taggcgtgtg caccttccgt ctgagacctg taaatggtta aaaaggaata
     4561 tataattacc ttttgaaaag tggtaaaaaa cgaatatatg tttttttttg gttttaatac
     4621 gctcttttgt gtgtatgaga ggaataaata agtgtgtgtg tgtgtgtgtg ttgttgttgt
     4681 tgaaaaaatg aaggacacga aagcgctcat taggggtggg gatacaccag gcataattat
     4741 ccgaaaatat tttggtgcac caaataagtg aacatactga cgaaatcaaa gagttggagg
     4801 atggataggg aggcctcatt ggcagtggta aattgatttg acttgaatat gataccctct
     4861 tactgtttca ttgccactac aagttggttt ccttcccctg caggcatgca agctagcttg
     4921 tattctatag tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta
     4981 gccgcgttct aacgacaata tgtacaagcc taattgtgta gcatctggct tactgaagca
     5041 gaccctatca tctctctcgt aaactgccgt cagagtcggt ttggttggac gaaccttctg
     5101 agtttctggt aacgccgtcc cgcacccgga aatggtcagc gaaccaatca gcagggtcat
     5161 cgctagccag atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     5221 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     5281 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     5341 gggcgccatc tccttgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     5401 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaatggt gcactctcag
     5461 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5521 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5581 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5641 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5701 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5761 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5821 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5881 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5941 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6001 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6061 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6121 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6181 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6241 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacagcatgg gggatcatgt
     6301 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6361 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6421 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6481 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6541 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6601 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6661 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6721 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6781 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6841 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6901 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6961 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7021 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7081 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7141 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7201 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7261 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7321 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7381 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7441 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7501 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7561 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7621 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7681 atgca
//
