LOCUS       sequence-310603-        6730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Long Fibronectin FRET Standard.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6730)
  AUTHORS   Duvall SW, Childers WS
  TITLE     Design of a Histidine Kinase FRET Sensor to Detect Complex Signal 
            Integration within Living Bacteria.
  JOURNAL   ACS Sens. 2020 Jun 26;5(6):1589-1596. doi: 
            10.1021/acssensors.0c00008. Epub 2020 Jun 16.
  PUBMED    32495620
REFERENCE   2  (bases 1 to 6730)
  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/acssensors.0c00008"; journalName: "ACS Sens"; date: 
            "2020-06-26- 26"; volume: "5"; issue: "6"; pages: "1589-1596"
COMMENT     Sequence Label: pSWD56
FEATURES             Location/Qualifiers
     source          1..6730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            27..136
                     /note="incP origin of transfer"
     primer_bind     288..304
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      305..351
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2893..2914)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3501..3522
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4117..4821
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="SKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMRI
                     KVIEGGPLPFAFDILGTSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGVV
                     TVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDIA
                     LKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVAK
                     YSNLGGGMDELYK"
     primer_bind     complement(4840..4856)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4867..4884)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4867..4883)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4876..4898)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5081..5669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5161..5180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5996..6655
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6062..6081)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ccccccccac cgcggtggcg gccgccccgg ccagcctcgc agagcaggat tcccgttgag
       61 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
      121 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
      181 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
      241 gaccccgaag cagggttatg cagcggaaaa gatccgtcga cggccgctct agaactagtg
      301 gatccaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      361 gtcgcattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
      421 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt ttttatataa
      481 gcttagcgct accggactca gatccagatc gaaatgtacc ggctgaactg gtggaagcac
      541 gttgagcctc gtcaaggact catgaagcgc gtccctgctc tcagtcgcct gctcgttacg
      601 cctgaggtgt caaaacaccg tctgttcatt tggctagacg ctagagttct tccggatcac
      661 aagctccagg tcgttacact cgatgacgac tgctcattcg gagttttaca ctcaaggttc
      721 cacgaggtgt gggctcttgc agcggggtcg tggcacggct caggcaatga tccgcgctac
      781 accatctcca ccaccttcga gaccttcccc ttcccggaag gcctgacgcc caacatagcc
      841 gccgtcgact atgagggcga cccgcgcgcc caggccatcg ccgcggcggc ggccgagctg
      901 aaccggctgc gggaagcctg gctcaatccg cccgacctgg tgcggatcga gcccgaggtc
      961 gtgcccggct atcccgaccg cgtgctgccc gtcagccccg aggcgggcgc cgagctgaag
     1021 aagcgcaccc tgaccaacct ctacaaccag cgcccggcct ggctggacat ggcccaccag
     1081 cgcctggacg ccgccgtcgc cgccgcctac ggctggcccg acggcctgac cgacgacgag
     1141 atcctggagc gcctgttcgc cctcaaccag gagcgcgcgg cggcggggcg gtaggttaga
     1201 cccctccaaa accccgacct ccccggcgaa ggccggggtc cagttacatc cggaacgctt
     1261 ggagtgggcc cgcctacggc ctcgaccaac ccgcagtcgg caaggggttc gatctggacc
     1321 ccggcattcg ccggggaggt cggcagagag gtcagaaccg cgtcttgcag ccctgcttca
     1381 gatgaacccc gcgcaggaag ccgcggcggg ccaggccggc gtcgacggcg gcgttcatct
     1441 ggcggtcggc gggggcagcg ccggtgactt cgcgcaccag gccccggaac gggatgatgc
     1501 tgttgatcag ggtcttgcca ccggcctcgg ccagcttgcc ggctcggttt tcggcgtact
     1561 cggcgccgac cttgtagtcg ggccccagcg ccgcgttcag cgccttcacc tcattggcca
     1621 gggcctggca ggacttcaga cccgcgaggc tgtaggggtc gtcaaaggcc ttggccagga
     1681 ccggcggcac ctcgcgcttg gacaggccga cgtcgcgcac cggctgggtg ccgatgtcga
     1741 cggccttctt gcccgcctcg ccccagcggc cggacggttc ctcgggcttg gccggcggct
     1801 tgacctgggc gtgggcgctg gaggccagca gcacggcggc ggctccggcc atgatcagca
     1861 gcttgttcgg catcaaaagc gccctcccga caaaacacgc cacgggaacg gcaagcctcg
     1921 ccgccggttc ccgcccgcaa gtccctcggc acgaactcta aggcctgcgc acgatccatc
     1981 gccaccacaa agcgtgtcat cccggccgaa gcgcagcgga gagccgggac ccagggccac
     2041 aagcacggcg cttcgtcacc ccctgggtcc cggatagcgc tccgcgcttc cgggatgaca
     2101 cgggtttaag cacttgggtg agaagcccta gttcggcgcg ccgcccgctt ccaggtcgaa
     2161 cgcggcgtcg aaggcgtcgt cgccaggaac cgtcccttcc agggcggcgt cgaacaggtc
     2221 ttcgagcgat gaccagatga aggccatctg ctccatgatc gtcgggacgc tggccggatc
     2281 gaagccgggg tagaggtctt ccaccagcca caggcccgtg ccgggatcga aggcgaagcg
     2341 gcagccgatc aggcggaact gggcgcgggc catggtctcg aacagggccg tcaggtcgcg
     2401 ggcggcgtcc aggtcgtcgt ggtccagcac caggcagcgg atctgcagcc agccgtggtc
     2461 gggcagcagg tagaaggcgc cctcgtcctg atcctcgccc gaaacctcca gcccccggtc
     2521 gatggcttcg acgacatagc cggccgcgcg gcaggtgtcg gtgagcgcgg ccagcagggc
     2581 ggcttcctgg tcaggggtca ggtcggtcat gggcaagagg tccaggtcgt ggtttgtcgg
     2641 cggcttctag catggaccgc ccgcgcccgt gaggccgagg atttcgcgct ggtcagacaa
     2701 cctacttgcc gtccccacat gttagcgcta ccaagtgccg acgaacgcgc gccgccgacg
     2761 gtgtcggcgc ttcagacgct cgagttttgg ggagacgacc atatgcctgc aggcgcctta
     2821 attaatatgc atggtacctt aagatctcga atgagcaagg gcgaggagct gttcaccggg
     2881 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtccgc
     2941 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc
     3001 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cgtggcctgc
     3061 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3121 ggctacgtcc aggagcgcac catctctttc aaggacgacg gtacctacaa gacccgcgcc
     3181 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3241 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag ccactacgtc
     3301 tatatcacgg ccgacaagca gaagaactgc atcaaggcta acttcaagat ccgccacaac
     3361 gttgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3421 ggccccgtgc tgctgcccga caaccactac ctgagccatc agtccaagct gagcaaagac
     3481 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cctggaaagc
     3541 ggcggcgaag atccgatggt gagcccgctg tcgcccccca ccaacctcca cctggaggcc
     3601 aatcccgata cgggcgtgct gacggtgtcc tgggagcgca gcacgacgcc cgacatcacc
     3661 gggtatcgta tcacgacgac gccgacgaac gggcagcagg ggaactcgct cgaagaagtc
     3721 gtgcacgccg accagtccag ctgcacgttc gacaatctct cgcccgggct ggagtacaac
     3781 gtctccgtct ataccgtgaa ggacgacaag gaatcggtcc cgatcagcga tacgatcatc
     3841 ccggcggtgc cgccccccac ggacctgcgg ttcacgaata tcggcccgga caccatgcgt
     3901 gtgacctggg cgccgccccc gtcgatcgac ctgacgaact tcctggtccg ttattccccg
     3961 gtgaagaacg aagaagacgt ggcggagctg agcatcagcc cgtcggacaa tgcggtggtc
     4021 ctcacgaatc tcctgccggg gaccgaatat gtcgtctcgg tctcctccgt gtacgagcag
     4081 cacgaatcca cgcccctgcg cgggcgccaa aagacctcta agggcgaaga gctgatcaag
     4141 gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca attcaaatgc
     4201 acaggtgaag gagaaggcag accgtacgag ggagtgcaaa ccatgaggat caaagtcatc
     4261 gagggaggac ccctgccatt tgcctttgac attcttggca cgtcgttcat gtatggcagc
     4321 cgtaccttta tcaagtaccc ggccgacatc cctgatttct ttaaacagtc ctttcctgag
     4381 ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac cgtcacgcag
     4441 gacaccagcc ttgaggatgg cgagctcgtc tacaacgtca aggtcagagg ggtaaacttt
     4501 ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa tacagagatg
     4561 atgtatccag cagatggtgg tctgagagga tacactgaca tcgcactgaa agttgatggt
     4621 ggtggccatc tgcactgcaa cttcgtgaca acttacaggt caaaaaagac cgtcgggaac
     4681 atcaagatgc ccggtgtcca tgccgttgat caccgcctgg aaaggatcga ggagagtgac
     4741 aatgaaacct acgtagtgca aagagaagtg gcagttgcca aatacagcaa ccttggtggt
     4801 ggcatggacg agctgtacaa gtaaagctgc agcccggggg atccactagt tctagagcgg
     4861 ccattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4921 taatcgcctt gcagcacatc cccctttcgc cagcggtatc agctcactca aaggcggtaa
     4981 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5041 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5101 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5161 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5221 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5281 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5341 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5401 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5461 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5521 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5581 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5641 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5701 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catcgaaccc cagagtcccg
     5761 gctagatcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     5821 gcggttttca tggcttgtta tgactgtttt tttggggtac cgtctatgcc tcgggcatcc
     5881 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     5941 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgaggaggta acgtcatgga
     6001 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     6061 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     6121 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     6181 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     6241 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     6301 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     6361 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     6421 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     6481 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     6541 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     6601 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaacctag
     6661 gcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     6721 gctttccccc
//
