LOCUS       sequence-360324-        4283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes PpmrD-mNeonGreen reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4283)
  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 4283)
  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: pKB233.3
FEATURES             Location/Qualifiers
     source          1..4283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(240..1190)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIMNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1238..1460
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(1952..2046)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2076..2867,2868..2936))
                     /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     2699..2718
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2937..3041)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3106..3124
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             3319..3330
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4056..4127
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4143..4170
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(4244..4261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgagcggtat cagctcactc aaaggcggta atctcgaggt tacattgtcg atctgttcat
       61 ggtgaacagc tttgaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta
      121 caccgttttc atctgtgcat atggacagtt ttccctttga tatgtaacgg tgaacagttg
      181 ttctactttt gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct
      241 cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc
      301 catgagaacg aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca
      361 aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta
      421 tgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg
      481 ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac
      541 gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa
      601 tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt
      661 tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt
      721 ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca
      781 aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa aagataaggc
      841 aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc
      901 cactggaaaa tctcaaagcc tttaaccaaa ggattcatga tttccacagt tctcgtcatc
      961 agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc
     1021 tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaata
     1081 gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca
     1141 tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat
     1201 tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacat
     1261 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
     1321 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
     1381 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
     1441 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
     1501 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
     1561 ctcgcaagct cgtgcaaatc gcttaatatt ccttttgtct ccgaccatca ggcacctgag
     1621 tcgctgcctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatgtggg
     1681 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
     1741 aaagcccgtc acgggcttct cagggcgttt tatggctggt ctgctatgtg gtgctatctg
     1801 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
     1861 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
     1921 gcttacccgt cttactgtcc ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     1981 aaccgagcgt tctgaacaaa tccagatgga gttctgatgt cattactgga tctatcaaca
     2041 ggagtccaag cgagctcgta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2101 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2161 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgtgat
     2221 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2281 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2341 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2401 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2461 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2521 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2581 tctcttactg tcatgccatc cgtaagatgc ttttctgtta ctggtgagta ctcaaccaag
     2641 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacggtat
     2701 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2761 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2821 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacggga
     2881 aggcaaaatg ccgcaaaaaa gtgaataagg gcgacactga aatgttgaat actcatactc
     2941 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3001 ttttaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc catggtgcca
     3061 cctgacgtct aataaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3121 aggcagaatt tttatcgctt ccgcgttccg tctactccct attacgctcc ggaatctccg
     3181 acgcctacgg ccgacgtcgg agtgagaact gtaaaaatta atgttaggtt aatatacaac
     3241 cattccatcg ctattgccgt tttgtttatc cgttgatgtg taacgtgtta tgaacaatca
     3301 gcgtgaaacg ggggcgctaa agaggagaaa tactagatgg tttcgaaagg tgaggaggat
     3361 aacatggcgt cgctgcctgc tacacatgaa ctgcatattt tcggcagcat caacggagtt
     3421 gatttcgaca tggtgggcca aggcaccggc aacccgaacg acggctatga agaattgaat
     3481 ttgaaatcta ccaagggcga cttgcaattt agtccgtgga ttctggtacc gcatattggt
     3541 tatggttttc atcagtatct gccctaccca gacggaatga gcccatttca ggcggctatg
     3601 gtggatggca gtggctatca ggtccatcgc accatgcaat tcgaggatgg ggcgagcctg
     3661 accgtgaatt atcggtacac ttacgaaggt tcacatatta aaggtgaagc acaagttaaa
     3721 ggcaccggct tcccggcgga tggcccagtg atgaccaaca gcctgacggc ggcggactgg
     3781 tgtcgtagca aaaagacgta tcccaatgat aaaacaatta tttctacctt taaatggagt
     3841 tacaccacgg gtaacggtaa acgctaccgt agcacggctc gcaccacata cacctttgcg
     3901 aaacccatgg ccgcgaacta tctgaaaaat cagccgatgt atgtgtttcg caaaacggaa
     3961 ctgaaacatt cgaaaactga actcaatttc aaagagtggc aaaaagcctt caccgacgta
     4021 atgggcatgg atgaactgta caaatgacca ggcatcaaat aaaacgaaag gctcagtcga
     4081 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     4141 ctggctcacc ttcgggtggg cctttctgcg tttatagcaa ggtgtcacca ccctgccctt
     4201 tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc
     4261 tgcgctcggt cgttcggctg cgg
//
