LOCUS       sequence-308582-        4726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High-copy plasmid to express the tVYN-Tm-Delta luminescent biosensor
            for in vitro purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4726)
  AUTHORS   Dippel AB, Anderson WA, Park JH, Yildiz FH, Hammond MC
  TITLE     Development of Ratiometric Bioluminescent Sensors for in Vivo 
            Detection of Bacterial Signaling.
  JOURNAL   ACS Chem Biol. 2020 Mar 25. doi: 10.1021/acschembio.9b00800.
  PUBMED    32186367
REFERENCE   2  (bases 1 to 4726)
  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: "ACS Chem 
            Biol. 2020 Mar 25. doi: 10.1021/acschembio.9b00800."
COMMENT     Sequence Label: pRSETb-tVYN-Tm-Delta
FEATURES             Location/Qualifiers
     source          1..4726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     845..864
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        845..863
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             895..917
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             937..954
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             958..990
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(994..1002,1003..1017)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1024..1026,1027..1029,1030..1542)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1069..1090)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1330..1349)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1677..1698
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2952..2970)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2966..3013
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3110..3565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3197..3216)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3407..3428
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3591..3695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3696..3764,3765..4556)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3914..3933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct cgatcccgcg
      841 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
      901 taactttaag aaggagatat acatatgcgg ggttctcatc atcatcatca tcatggtatg
      961 gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
     1021 ccgatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1081 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1141 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     1201 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     1261 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1321 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1381 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1441 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     1501 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1561 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1621 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1681 gtcctgctgg agttcgtgac cgccggtaca ggttataccg aactggtgaa cgcgaaagat
     1741 gtgattcgcc cgggccagaa cgtgattgtg gaagtgagcg cgccggaaga tctggaaggc
     1801 cagtataaaa gcagcgtgca tgatgtggat tttgaaaaac gcgtgctgac cctgagcatg
     1861 ccaagttttc gtggtcgtct ggtgccactg ccacgtggta cccgttgcac ggtgatgatt
     1921 ctggatagca gcgcgattta tgtgtttcgc accagcgtgc tggaaagcgg ccgcgatgaa
     1981 gatggctttc cggtgaccaa agtgccgttt ccgggccgcc tgcgcaaaat tcagcgccgc
     2041 cgctttaaac gcattaaaat ttttctggaa ggcacctatc gcgtggcgag ccgcgatgaa
     2101 ccgccgaaac gctttgtgac ccgcgatttt agcgcgggcg gcatgctgat ggtggtggaa
     2161 gatattctga ccccggaaca gattatttat gtgaccctgg atctggatga agatctgaaa
     2221 ctgaaagatc atccagcgcg tgtggttcgt gaagccggtg ccctggaaac cggtgaacgc
     2281 atgtatggcg tggaatttct gaacgtgccg ccggcgctgg aacgcaaact ggtgagcttt
     2341 gtgtttaaaa aagaaattga aatgcgcaac aaagaaggtg gtaccctcga agatttcgtt
     2401 ggggactggc gacagacagc cggctacaac ctggaccaag tccttgaaca gggaggtgtg
     2461 tccagtttgt ttcagaatct cggggtgtcc gtaactccga tccaaaggat tgtcctgagc
     2521 ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcggc
     2581 gaccaaatgg gccagatcga aaaaattttt aaggtggtgt accctgtgga tgatcatcac
     2641 tttaaggtga tcctgcacta tggcacactg gtaatcgacg gggttacgcc gaacatgatc
     2701 gactatttcg gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta
     2761 acagggaccc tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa ccccgacggc
     2821 tccctgctgt tccgagtaac catcaacgga gtgaccggct ggcggctgtg cgaacgcatt
     2881 ctggcgtaag aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     2941 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3001 gaggggtttt ttgctgaaag gaggaactat atccggatct ggcgtaatag cgaagaggcc
     3061 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3121 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3181 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3241 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3301 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3361 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3421 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3481 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3541 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3601 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3661 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3721 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3781 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3841 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3901 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3961 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4021 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4081 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4141 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4201 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4261 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4321 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4381 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4441 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4501 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4561 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
