LOCUS       sequence-452445-        4089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP inducible Lysin KU1 Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4089)
  AUTHORS   Baumer ZT, Newton MS, Lofstrand L, Carpio Paucar GN, Farny NG, 
            Whitehead TA
  TITLE     Engineered Stop and Go T7 RNA Polymerases.
  JOURNAL   ACS Synth Biol. 2024 Dec 20;13(12):4165-4174. doi: 
            10.1021/acssynbio.4c00627. Epub 2024 Nov 28.
  PUBMED    39610115
REFERENCE   2  (bases 1 to 4089)
  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"; doi: 
            "10.1021/acssynbio.4c00627"; journalName: "ACS Synth Biol"; date: 
            "2024-12-20- 20"; volume: "13"; issue: "12"; pages: "4165-4174"
COMMENT     Sequence Label: pZB641_pT7_Lysin-KU1
FEATURES             Location/Qualifiers
     source          1..4089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(578..672)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(703..1497)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(814..833)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1424..1443
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      1652..1738
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1830..1857
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1935..1953
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1996..2015
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1996..2014
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2051..2062
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2331..2417
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2955..3905)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      join(3953..4089,1..86)
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga
       61 atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg
      121 atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt
      181 agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca
      241 cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa
      301 tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat
      361 acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc
      421 tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg
      481 gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat
      541 caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata aaaaacgccc
      601 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
      661 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag
      721 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
      781 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
      841 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
      901 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
      961 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     1021 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     1081 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     1141 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     1201 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     1261 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     1321 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     1381 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     1441 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     1501 aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg
     1561 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     1621 agctggcaat tccgacgtca ccggtaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1681 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1741 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     1801 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     1861 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1921 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaaacg cgtgtacacc
     1981 cgggcggccg caaattaata cgactcacta tagggagatc tccatacccg tttttttggg
     2041 ctagcgaatt aaagaggaga aaggtcatgg gtcagaaagc atttaaacat aaggaaaacc
     2101 tatgttccga ttcacagaga tcgaaaagac tctatgtatg gatcgcactc gcgattgtgc
     2161 tgtccgattt cacgtctatc ttcagtcatt ggatttgggg tcttctgatc ctcattctcc
     2221 ggactttgat ggacttgcct accttcgtga tgaatgttta ataagcttgg ctgttttggc
     2281 ggatgagagc gcgcttaatg cgccgctaca gggcgcgtct ctagaggcat caaataaaac
     2341 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2401 tcctgagtag gacaaatccg ccgccctaga cctagggtac gggttttgct gcccgcaaac
     2461 gggctgttct ggtgttgcta gtttgttatc agaatcgcag atccggcttc agccggtttg
     2521 ccggctgaaa gcgctatttc ttccagaatt gccatgattt tttccccacg ggaggcgtca
     2581 ctggctcccg tgttgtcggc agctttgatt cgataagcag catcgcctgt ttcaggctgt
     2641 ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg ttcaatttca tgttctagtt
     2701 gctttgtttt actggtttca cctgttctat taggtgttac atgctgttca tctgttacat
     2761 tgtcgatctg ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg
     2821 tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt
     2881 aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa
     2941 gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt
     3001 tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc
     3061 aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt
     3121 tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca
     3181 ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca
     3241 acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg
     3301 ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac
     3361 tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata
     3421 tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata
     3481 gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac
     3541 tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac
     3601 ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc
     3661 acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc attttcccta
     3721 ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg ttctttcctt
     3781 gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca
     3841 tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca
     3901 gacatacatc tcaattggtc taggtgattt taatcactat accaattgag atgggctagt
     3961 caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt ctgctagacc
     4021 tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag acctttgtgt
     4081 gtttttttt
//
