LOCUS       sequence-336782-        4286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes PvirK-mNeonGreen reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4286)
  AUTHORS   Brink KR, Mu AM, Hoang KV, Groszman K, Gunn JS, Tabor JJ
  TITLE     High-throughput discovery of peptide activators of a bacterial 
            sensor kinase
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 4286)
  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: "bioRxiv"
COMMENT     Sequence Label: pKB233
FEATURES             Location/Qualifiers
     source          1..4286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      493..715
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             763..1713
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIMNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     primer_bind     1975..1992
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2066..2093)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2109..2180)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2906..2917
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(3115..3133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3198..3302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3303..3371,3372..4163)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFSVALIHFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLYRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3521..3540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      join(4193..4286,1)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 tccaagcact agggacagta agacgggtaa gcctgttgat gataccgctg ccttactggg
       61 tgcattagcc agtctgaatg acctgtcacg ggataatccg aagtggtcag actggaaaat
      121 cagagggcag gaactgctga acagcaaaaa gtcagatagc accacatagc agaccagcca
      181 taaaacgccc tgagaagccc gtgacgggct tttcttgtat tatgggtagt ttccttgcat
      241 gaatccataa aaggcgcctg tagtgccatt tacccacatt cactgccaga gccgtgagcg
      301 cagcgaactg aatgtcacga aaaaggcagc gactcaggtg cctgatggtc ggagacaaaa
      361 ggaatattaa gcgatttgca cgagcttgcg agggtgctac ttaagccttt agggttttaa
      421 ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact
      481 gactaaagta gtgagttata cacagggctg ggatctattc tttttatctt tttttattct
      541 ttctttattc tataaattat aaccacttga atataaacaa aaaaaacaca caaaggtcta
      601 gcggaattta cagagggtct agcagaattt acaagttttc cagcaaaggt ctagcagaat
      661 ttacagatac ccacaactca aaggaaaagg acatgtaatt atcattgact agcccatctc
      721 aattggtata gtgattaaaa tcacctagac caattgagat gtatgtctga attagttgtt
      781 ttcaaagcaa atgaactagc gattagtcgc tatgacttaa cggagcatga aaccaagcta
      841 attttatgct gtgtggcact actcaacccc actattgaaa accctacaag gaaagaacgg
      901 acggtatcgt tcacttataa ccaatacgct cagatgatga acatcagtag ggaaaatgct
      961 tatggtgtat tagctaaagc aaccagagag ctgatgacga gaactgtgga aatcatgaat
     1021 cctttggtta aaggctttga gattttccag tggacaaact atgccaagtt ctcaagcgaa
     1081 aaattagaat tagtttttag tgaagagata ttgccttatc ttttccagtt aaaaaaattc
     1141 ataaaatata atctggaaca tgttaagtct tttgaaaaca aatactctat gaggatttat
     1201 gagtggttat taaaagaact aacacaaaag aaaactcaca aggcaaatat agagattagc
     1261 cttgatgaat ttaagttcat gttaatgctt gaaaataact accatgagtt taaaaggctt
     1321 aaccaatggg ttttgaaacc aataagtaaa gatttaaaca cttacagcaa tatgaaattg
     1381 gtggttgata agcgaggccg cccgactgat acgttgattt tccaagttga actagataga
     1441 caaatggatc tcgtaaccga acttgagaac aaccagataa aaatgaatgg tgacaaaata
     1501 ccaacaacca ttacatcaga ttcctaccta cataacggac taagaaaaac actacacgat
     1561 gctttaactg caaaaattca gctcaccagt tttgaggcaa aatttttgag tgacatgcaa
     1621 agtaagtatg atctcaatgg ttcgttctca tggctcacgc aaaaacaacg aaccacacta
     1681 gagaacatac tggctaaata cggaaggatc tgaggttctt atggctcttg tatctatcag
     1741 tgaagcatca agactaacaa acaaaagtag aacaactgtt caccgttaca tatcaaaggg
     1801 aaaactgtcc atatgcacag atgaaaacgg tgtaaaaaag atagatacat cagagctttt
     1861 acgagttttt ggtgcattca aagctgttca ccatgaacag atcgacaatg taacctcgag
     1921 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     1981 tcagtgagcg aggaagcctg cataacgcga agtaatcttt tcggttttaa agaaaaaggg
     2041 cagggtggtg acaccttgct ataaacgcag aaaggcccac ccgaaggtga gccagtgtga
     2101 ctctagtaga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac
     2161 tgagcctttc gttttatttg atgcctggtc atttgtacag ttcatccatg cccattacgt
     2221 cggtgaaggc tttttgccac tctttgaaat tgagttcagt tttcgaatgt ttcagttccg
     2281 ttttgcgaaa cacatacatc ggctgatttt tcagatagtt cgcggccatg ggtttcgcaa
     2341 aggtgtatgt ggtgcgagcc gtgctacggt agcgtttacc gttacccgtg gtgtaactcc
     2401 atttaaaggt agaaataatt gttttatcat tgggatacgt ctttttgcta cgacaccagt
     2461 ccgccgccgt caggctgttg gtcatcactg ggccatccgc cgggaagccg gtgcctttaa
     2521 cttgtgcttc acctttaata tgtgaacctt cgtaagtgta ccgataattc acggtcaggc
     2581 tcgccccatc ctcgaattgc atggtgcgat ggacctgata gccactgcca tccaccatag
     2641 ccgcctgaaa tgggctcatt ccgtctgggt agggcagata ctgatgaaaa ccataaccaa
     2701 tatgcggtac cagaatccac ggactaaatt gcaagtcgcc cttggtagat ttcaaattca
     2761 attcttcata gccgtcgttc gggttgccgg tgccttggcc caccatgtcg aaatcaactc
     2821 cgttgatgct gccgaaaata tgcagttcat gtgtagcagg cagcgacgcc atgttatcct
     2881 cctcaccttt cgaaaccatc tagtatttct cctctttact acggctctac tactcgatag
     2941 acttataacc gagacggtag cctgattgag ttaaacgtaa aggcaacgaa gaaggtctgt
     3001 acagacgatg ttgttaaaca gtttatcaat ggcggtatcc gacgtcggcc gtaggcgtcg
     3061 gagattccgg agcgtaatag ggagtagacg gaacgcggaa gcgataaaaa ttctgcctcg
     3121 tgatacgcct atttttatag gttaatgtca tgataataat ggtttattag acgtcaggtg
     3181 gcaccatggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattaaaata
     3241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3301 gtatgagtat tcaacatttc agtgtcgccc ttattcactt ttttgcggca ttttgccttc
     3361 ccgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3421 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3481 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3541 accgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3601 tggttgagta ctcaccagta acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3661 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3721 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3781 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3841 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3901 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3961 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat
     4021 cacgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4081 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4141 cctcactgat taagcattgg taactgtcag accaagttta cgagctcgct tggactcctg
     4201 ttgatagatc cagtaatgac atcagaactc catctggatt tgttcagaac gctcggttgc
     4261 cgccgggcgt tttttattgg tgagaa
//
