LOCUS       sequence-11283-e        8297 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 8297)
  TITLE     George Cross Lab Plasmids
REFERENCE   2  (bases 1 to 8297)
  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: pLEW100v5-HYG
FEATURES             Location/Qualifiers
     source          1..8297
                     /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(1301..2326)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(1301..2326)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(2469..2488)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2470..2488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2765..2783
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2786..2804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3000..4652
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKRLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(3066..3084)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(5536..5554)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5537..5554)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5844..5863)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6073..6092)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6252..6270)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6338..6442
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6443..6511,6512..7303)
                     /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(6661..6680)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7474..8062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7963..7982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8216..8233
                     /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 tttgcagaat actgcataga
     1021 taacaaacgc atcaaacaca actggtggca ctatttcaat catgtcgaca caccaagacg
     1081 agaaaatttg agttaaacaa gatgcctaca accctatatt tattatcttc aagtgatctt
     1141 ctcttctctt gtgcgctgta cgtaaatgtg ttgcaaataa ggcatagggc tgagtacagg
     1201 caccacacgc gaggaaaatc ataagcacta gccgtgtcaa ctcacagcga ctacagaaca
     1261 attttggcca cacaacccgg tgttaggatc tccgaggcct ctattccttt gccctcggac
     1321 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     1381 cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     1441 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     1501 gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa
     1561 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     1621 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     1681 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     1741 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     1801 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     1861 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     1921 atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg
     1981 cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     2041 gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt
     2101 ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     2161 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     2221 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     2281 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttcatcact agttctagag
     2341 cttattttat ggcagcaacg agaccttacg taaagcaaaa ctaataagag cggagactgc
     2401 aatgcagagt aaaaaaaaaa aaacatgaat ttcagaagac cttgctgtgc ccccggtacg
     2461 ggagatctcc ctatagtgag tcgtattaat caggtaccta gctttccacc cagcgcgggt
     2521 gcattctggc tcttatatat acttattgtc atgacagagt atattgtact gtgttgataa
     2581 gggacgggta actgtattga agagccgatg cttttgacat gttagatata atatgtttta
     2641 ttgtaaagtc aatacaacac acaataggat aataatgata aagttaaaaa agtatatata
     2701 gtaatagaaa tatatcttat ataggaaaga ttaagcagta aaagtagcgc ttacggcgta
     2761 cggttcccta tcagtgatag agatctccct atcagtgata gagatccctg agtactgagt
     2821 ttaacatgtt ctcgtcccgg gctgcacgcg ccttcgagtt ttttttcctt ttccccattt
     2881 ttttcaactt gaagacttca attacaccaa aaagtaaaat tcacaagctt ggaattcctt
     2941 tgtgttacat tcttgaatgt cgctcgcagt gacattagca ttccggtact gttggtaaaa
     3001 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatcctcta gaggatggaa
     3061 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
     3121 cttttacaga tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg
     3181 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
     3241 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
     3301 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgaacattt
     3361 cgcagcctac cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
     3421 aaaaattacc aataatccag aaaattatta tcatggattc taaaacggat taccagggat
     3481 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
     3541 ttgtaccaga gtcctttgat cgtgacaaaa caattgcact gataatgaat tcctctggat
     3601 ctactgggtt acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc
     3661 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
     3721 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
     3781 gagtcgtctt aatgtataga tttgaagaag agctgttttt acgatccctt caggattaca
     3841 aaattcaaag tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga
     3901 ttgacaaata cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga
     3961 aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg
     4021 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
     4081 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
     4141 cgctgggcgt taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt
     4201 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
     4261 gagacatagc ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt
     4321 taattaaata caaaggatat caggtggccc ccgctgaatt ggaatcgata ttgttacaac
     4381 accccaacat cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc
     4441 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
     4501 acgtcgccag tcaagtaaca accgcgaaaa ggttgcgcgg aggagttgtg tttgtggacg
     4561 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
     4621 aggccaagaa gggcggaaag tccaaattgt aaaatgtaac tgtattcagc gatgacgaaa
     4681 ttcttagcta ttgtaatatt atatgcaaat tgatgaatgg taattttgta attgtgggtc
     4741 actgtactat tttaacgaat aataaaatca ggtataggta actaaaaagg aattcgagct
     4801 cggatcctgc ccatttagtt ggcttttccc ttgtctcgtg tcttttccgt ggaaaggttc
     4861 ccggagtaat ctgatggcac agcagggagg tgcgcctgca ggttggttag gaagggggga
     4921 tgatgtaaaa gaagaaaatg gggggatatc ttatgtttaa gaggataaat aatgtgaagg
     4981 ggctttatat gcttgcttgg ttgtctcgtt ggtgcatggg gaatctgcat gtttgctttg
     5041 gagcacgcgt ggtacatcaa tgggatcata tcttgctgcc tcccgcagtc accgtgcgga
     5101 gctgccggtg ccccgtttct cataggcgtg tgcaccttcc gtctgagacc tgtaaatggt
     5161 taaaaaggaa tatataatta ccttttgaaa agtggtaaaa aacgaatata tgtttttttt
     5221 tggttttaat acgctctttt gtgtgtatga gaggaataaa taagtgtgtg tgtgtgtgtg
     5281 tgttgttgtt gttgaaaaaa tgaaggacac gaaagcgctc attaggggtg gggatacacc
     5341 aggcataatt atccgaaaat attttggtgc accaaataag tgaacatact gacgaaatca
     5401 aagagttgga ggatggatag ggaggcctca ttggcagtgg taaattgatt tgacttgaat
     5461 atgataccct cttactgttt cattgccact acaagttggt ttccttcccc tgcaggcatg
     5521 caagctagct tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat
     5581 gtattaattg tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg
     5641 cttactgaag cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg
     5701 acgaaccttc tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat
     5761 cagcagggtc atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg
     5821 cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc
     5881 tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
     5941 cgggggactg ttgggcgcca tctccttgca ccattccttg cggcggcggt gctcaacggc
     6001 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaatg
     6061 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6121 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6181 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6241 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6301 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6361 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6421 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6481 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6541 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6601 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6661 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6721 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6781 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6841 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacagcat
     6901 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6961 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7021 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7081 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7141 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7201 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7261 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7321 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7381 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7441 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7501 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7561 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7621 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7681 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7741 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7801 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7861 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7921 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7981 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8041 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8101 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8161 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     8221 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     8281 gccgattcat taatgca
//
