LOCUS       sequence-11280-e        6641 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 6641)
  TITLE     George Cross Lab Plasmids
REFERENCE   2  (bases 1 to 6641)
  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: pLEW82v4
FEATURES             Location/Qualifiers
     source          1..6641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        720..738
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    745..763
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    771..789
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    797..815
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             983..2635
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1049..1067)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             3112..3486
                     /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"
     terminator      3889..3936
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4022..4069
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4186..4205)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4416..4435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4595..4613)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4681..4785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4786..4854,4855..5646)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5004..5023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5817..6405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6306..6325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6559..6576
                     /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 tagggaagcg acgcttgccc
      481 tagtcccacg agtaaccaag actccaaaag cctttctggc acagagagcg agccgaaatg
      541 gaaaagagaa acaatgcctg cactaacact actgagcgat tcgcctcgcc gcggaggacc
      601 gaatactaat aacgacactt gcggtcaaaa agtagaagaa caaatgctca acgatgagtg
      661 aatcaggtta gggtagttgg aaaattatac agaatgtctt tggcaacaca ccggctgatt
      721 aatacgactc actataggga gatctcccta tcagtgatag agacatccgc tccctatcag
      781 tgatagagac ggtagctccc tatcagtgat agagatctcc cgggctgcac gcgccttcga
      841 gttttttttc cttttcccca tttttttcaa cttgaagact tcaattacac caaaaagtaa
      901 aattcacaag cttggaattc ctttgtgtta cattcttgaa tgtcgctcgc agtgacatta
      961 gcattccggt actgttggta aaatggaaga cgccaaaaac ataaagaaag gcccggcgcc
     1021 attctatcct ctagaggatg gaaccgctgg agagcaactg cataaggcta tgaagagata
     1081 cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtga acatcacgta
     1141 cgcggaatac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa
     1201 tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt
     1261 gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga
     1321 attgctcaac agtatgaaca tttcgcagcc taccgtagtg tttgtttcca aaaaggggtt
     1381 gcaaaaaatt ttgaacgtgc aaaaaaaatt accaataatc cagaaaatta ttatcatgga
     1441 ttctaaaacg gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc
     1501 tcccggtttt aatgaatacg attttgtacc agagtccttt gatcgtgaca aaacaattgc
     1561 actgataatg aattcctctg gatctactgg gttacctaag ggtgtggccc ttccgcatag
     1621 aactgcctgc gtcagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc
     1681 ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact
     1741 cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt
     1801 tttacgatcc cttcaggatt acaaaattca aagtgcgttg ctagtaccaa ccctattttc
     1861 attcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc
     1921 ttctgggggc gcacctcttt cgaaagaagt cggggaagcg gttgcaaaac gcttccatct
     1981 tccagggata cgacaaggat atgggctcac tgagactaca tcagctattc tgattacacc
     2041 cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt
     2101 tgtggatctg gataccggga aaacgctggg cgttaatcag agaggcgaat tatgtgtcag
     2161 aggacctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga
     2221 caaggatgga tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt
     2281 catagttgac cgcttgaagt ctttaattaa atacaaagga tatcaggtgg cccccgctga
     2341 attggaatcg atattgttac aacaccccaa catcttcgac gcgggcgtgg caggtcttcc
     2401 cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat
     2461 gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg
     2521 cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag
     2581 aaaaatcaga gagatcctca taaaggccaa gaagggcgga aagtccaaat tgtaaaatgt
     2641 aactgtattc agcgatgacg aaattcttag ctattgtaat attatatgca aattgatgaa
     2701 tggtaatttt gtaattgtgg gtcactgtac tattttaacg aataataaaa tcaggtatag
     2761 gtaactaaaa aggaattcga gctcggatcc aagcttctag actcgagcat taatacgtct
     2821 cactataggg agaggatcct gcccatttag ttggcttttc ccttgtctcg tgtcttttcc
     2881 gtggaaaggt tcccggagta atctgatggc acagcaggga ggtgcgcctg caggttggtt
     2941 aggaaggggg gatgatgtaa aagaagaaaa tggggggatt cgagcccggg cacagcaagg
     3001 tcttctgaaa ttcatgtttt tttttttttt actctgcatt gcagtctccg ctcttattta
     3061 gttttgcttt acgtaaggtc tcgttgctgc cataaaataa gctatatcac catggccaag
     3121 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
     3181 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
     3241 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
     3301 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
     3361 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
     3421 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
     3481 gactgaccga cgccgaccaa caccgccggt ccgacggcgg cccacgggtc ccaggcctcg
     3541 gagatcctaa caccgggttg tgtggccaaa attgttctgt agtcgctgtg agttgacacg
     3601 gctagtgctt atgattttcc tcgcgtgtgg tgcctgtact cagccctatg ccttatttgc
     3661 aacacattta cgtacagcgc acaagagaag agaagatcac ttgaagataa taaatatagg
     3721 gttgtaggca tcttgtttaa ctcaaatttt ctcgtcttgg tgtgtcgaca tgattgaaat
     3781 agtgccacca gttgtgtttg atgcgtttgt tatctatgca gtattctgca ggctgctaac
     3841 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     3901 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     3961 tagggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     4021 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     4081 aactatatcc ggatagggcc cctgcaggca tgcaagctag ccagatcctc tacgccggac
     4141 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     4201 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     4261 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg caccattcct
     4321 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     4381 gcataaggga gagcgtcgat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4441 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4501 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4561 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4621 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4681 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4741 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4801 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4861 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4921 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4981 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5041 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5101 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5161 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5221 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5281 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5341 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5401 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5461 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5521 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5581 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5641 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5701 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5761 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5941 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6481 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6541 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6601 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca g
//
