LOCUS       sequence-503486-        4593 bp    DNA     circular SYN 19-MAR-2026
DEFINITION  Opto-CheY V416T expressed using the proA promoter and T7 RBS variant
            GAAGGAGgT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4593)
  AUTHORS   Vrabioiu AM, Hosu BG, Samuel ADT
  TITLE     The dynamic response of the bacterial flagellar motor to its direct 
            intracellular input signal.
  JOURNAL   Proc Natl Acad Sci U S A. 2026 Mar 10;123(10):e2516278123. doi: 
            10.1073/pnas.2516278123. Epub 2026 Mar 3.
  PUBMED    41774795
REFERENCE   2  (bases 1 to 4593)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2026-03-10"; volume: "123(10):e2516278123. 
            doi"; pages: " 10.1073/pnas.2516278123. Epub 2026 Mar 3"
COMMENT     Sequence Label: pA-Opto_V416T
FEATURES             Location/Qualifiers
     source          1..4593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..135
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1276..1362
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1900..2850)
                     /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      2898..3120
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3612..3706)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3737..4531)
                     /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(3848..3867)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4458..4477
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
       61 cagctggcaa ttccgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
      121 ggcgtatcac gaggcccttt cgtcttcacc acagctaaca ccacgtcgtc cctatctgct
      181 gccctaggtc tatgagtggt tgctggataa ctttacgggc atgcataagg ctcgtaggct
      241 atattcaggg agaccacaac ggtttccctc tacaaataat tttgtttaac ttttctagat
      301 ttaagaagga ggtatacata tggcggataa agaacttaaa tttttggttg tggatgactt
      361 ttccaccatg cgacgcatag tgcgtaacct gctgaaagag ctgggattca ataatgttga
      421 ggaagcggaa gatggcgtcg acgctctcaa taagttgcag gcaggcggtt atggatttgt
      481 tatctccgac tggaacatgc ccaatatgga tggcctggaa ttgctgaaaa caattcgtgc
      541 ggatggcgcg atgtcggcat tgccagtgtt aatggtgact gcagaagcga agaaagagaa
      601 catcattgct gcggcgcaag cgggggccag tggctatgtg gtgaagccat ttaccgccgc
      661 gacgctggag gaaaaactca acaaaatctt tgagaaagac gcggccgagc gtgagggcgt
      721 aatgttaatt aagaagaccg cagaaaacat tgatgaagct gctaaagagt tagggggtgg
      781 ctcaggcggg tctgggggcg gtttggccac cacgttggag cgcatcgaaa agaatttcac
      841 tatcactgac ccacgcttac cagacaaccc cattatcttt gctagtgatt cttttttgca
      901 acttacagag tattctcgtg aagaaattct tgggcgtaac tgtcgtttct tgcagggtcc
      961 tgagacggat cgcgcaacag ttcgtaagat tcgcgatgct atcgacaatc aaactgaggt
     1021 caccgtgcaa ctgatcaatt atactaaatc cgggaagaag ttttggaact tgtttcatct
     1081 tcagccaatg cgtgatcaaa agggagacgt gcaatacttt atcggcgttc aactggacgg
     1141 gactgagcat gtacgtgcaa aggaggctgc agcaaaagaa gctgcagcga aaggtggcga
     1201 tagcattctg agccaggcgg aaattgatgc gctgctgaat ggctgaacct gcaggcatgc
     1261 aagctctaga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1321 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag
     1381 ggtacgggtt ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat
     1441 cgcagatccg gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat
     1501 gattttttcc ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata
     1561 agcagcatcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc
     1621 aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt
     1681 gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca
     1741 ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat
     1801 atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt
     1861 cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc
     1921 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     1981 atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     2041 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt
     2101 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     2161 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     2221 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     2281 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     2341 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     2401 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     2461 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     2521 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     2581 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     2641 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     2701 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     2761 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     2821 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     2881 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     2941 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     3001 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     3061 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     3121 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     3181 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     3241 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     3301 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     3361 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     3421 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     3481 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     3541 ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc
     3601 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     3661 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg
     3721 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     3781 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     3841 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     3901 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     3961 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     4021 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     4081 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4141 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     4201 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     4261 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     4321 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     4381 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     4441 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4501 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     4561 cagatcttga tcccctgcgc catcagatcc ttg
//
