LOCUS       sequence-169038-        5676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bxb1/TP901 temporal logic gate with mKate2 and sfGFP on low-copy 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  AUTHORS   Hsiao V, Hori Y, Rothemund PW, Murray RM
  TITLE     A population-based temporal logic gate for timing and recording 
            chemical events.
  JOURNAL   Mol Syst Biol. 2016 May 17;12(5):869. doi: 10.15252/msb.20156663.
  PUBMED    27193783
REFERENCE   2  (bases 1 to 5676)
  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: "Mol Syst 
            Biol."; date: "2016-05-17"; volume: "12(5):869. doi"; pages: " 
            10.15252/msb.20156663"
COMMENT     Sequence Label: pVHed02
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(100..129)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    176..197
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      complement(1126..1153)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1169..1240)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1249..1298
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    1410..1457
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     misc_RNA        1464..1513
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             1607..1615
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1622..2332
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGTKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELY"
     terminator      2358..2429
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2445..2472
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2479..2499
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2535..2566
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      2691..2785
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3277..3499
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3547..4497
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNCAKFS
                     SEKLELVFSEGILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQFTSFEAKF
                     LGDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             join(4786..4854,4855..5646)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITTSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg tttgccacct
       61 gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc gaagaaaggc ccacccgtga
      121 aggtgagcca gtgagttgat tgctacgtaa ttagttagtt agcccttagt gactcgaatt
      181 cgcggccgct tctagagtta tcaacgatgt cctaatttcg acggaaggtc acaataacgg
      241 gctacggcta cttcatgttg ttcaacgtag gtttctttat cagcttcttt aatacgttct
      301 aagcgacgat caacatagta tactccaggc attttaaggt ttttagcggg ttttttggaa
      361 cgataggttg ttttcaggtt gcagattaaa tgaccgcctc caactaattt cagtgccatg
      421 tctgcgcgtc cttctaagcc accgtccgca ggatataagg tttctgttga tgcttcccaa
      481 ccaagagttt ttttttgcat tacgggtccg ttactcggga aatttacgcc acggatttta
      541 acattgtaaa tcagacagcc atcttgtaaa ctcgtatctt gagttgctgt aagtacaccg
      601 ccatcttcat aagttgttac acgttcccag gtaaagcctt caggaaagga ctgtttaaaa
      661 aagtcaggaa tgccttgggt gtgattgata aaagttttag acccgtacat aaagctcgtt
      721 gcgagaatgt caaacgcaaa gggcaggggt ccaccttcaa ctgctttgat gcgcattgtt
      781 tgtgtgcctt catatggttt accttcacct tcggatgtgc atttgaaatg atgattgttg
      841 acagtacctt ccatatataa tttcatgtgc atattttctt taattaattc tgacattaga
      901 aagtctcctg tgcatgatta agatgtttca gtacgaaaat tgctttcatt gttgatctcc
      961 tttttaagtg aacttgggcc cttaaacaaa attatttgta gaggataccg tttcggactt
     1021 ttgtactctt caggtgagac acacatccca ctacagcatc taagcttatg ccaacacaat
     1081 taacatctca atcaaggtaa atgctttttg ctttttttgc ataaacgcag aaaggcccac
     1141 ccgaaggtga gccagtgtga ctctagtaga gagcgttcac cgacaaacaa cagataaaac
     1201 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctgggc ccggatgatc
     1261 ctgacgacgg agaccgccgt cgtcgacaag ccggccgagc tagcaaaagg agttttttag
     1321 ttaccttaat tgaaataaac gaaataaaaa ctcgcaagct ttccacacat tataggtaca
     1381 aaaagatgcg aaagcaaata aattttttgg gtttgtaccg tacaccactg agaccgcggt
     1441 ggttgaccag acaaaccacg aagctgtcac cggatgtgct ttccggtctg atgagtccgt
     1501 gaggacgaaa cagcctctac aaataatttt gtttaagggc ccaagttcac ttaaaaagga
     1561 gatcaacaat gaaagcaatt ttcgtactga aacatcttaa tcatgctaag gaggttttct
     1621 aatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
     1681 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
     1741 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt ggccgactct
     1801 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
     1861 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
     1921 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg atactctggt
     1981 aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc tgggccataa
     2041 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
     2101 cactaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
     2161 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
     2221 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
     2281 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acagatgatg
     2341 atactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcccttcgtt
     2401 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     2461 ggcctttctg cgtttatata ctagtagcgg ccgctgcagg agtcactaag ggttagttag
     2521 ttagattagc agaaagtcaa aagcctccga ccggaggctt ttgactaaaa cttcccttgg
     2581 ggttatcatt ggggctcact caaaggcggt aatcagataa aaaaaatcct tagctttcgc
     2641 tgaggatgat ttctgctaga gctgtcagac caagtttacg agctcgcttg gactcctgtt
     2701 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     2761 ccgggcgttt tttattggtg agaatccaag cactagggac agtaagacgg gtaagcctgt
     2821 tgatgatacc gctgccttac tgggtgcatt agccagtctg aatgacctgt cacggggtaa
     2881 tccgaagtgg ccagactgga aaatcagagg gcaggaactg ctgaacagca aaaagtcaga
     2941 tagcaccgca tagcagaccc gccataaaac gccctgagaa gcccgtgacg ggcttttctt
     3001 gtattatggg tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc catttacccc
     3061 cattcactgc cagagccgtg agtgcagcga actgaatgtc acgaaaaaga cggcgactca
     3121 ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg
     3181 ctacttaagc ctttagggtt ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac
     3241 atccttttgt aatactgcgg aactgactaa agtagtgagt tatacacagg gctgggatct
     3301 attcttttta tcttttttta ttctttcttt attctataaa ttataaccac ttgaatataa
     3361 acaaaaaaaa cacacaaagg tctagcggaa tttacagagg gtctagcaga atttacaagt
     3421 tttccagcaa aggtctagca gaatttacag atacccacaa ctcaaaggaa aaggacatgt
     3481 aattatcatt gactagccca tctcaattgg tatagtgatt aaaatcacct agaccaattg
     3541 agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac tagcgattag tcgctatgac
     3601 ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg cactactcaa ccccacgatt
     3661 gaaaacccta caaggaaaga acggacggta tcgttcactt ataaccaata cgctcagatg
     3721 atgaacatca gtagggaaaa tgcttatggt gtattagcta aagcaaccag agagctgatg
     3781 acgagaactg tggaaatcag gaatcctttg gttaaaggct ttgagatttt ccagtggaca
     3841 aactgtgcca agttctcaag cgaaaaatta gaattagttt ttagtgaagg gatattgcct
     3901 tatcttttcc agttaaaaaa attcataaaa tataatctgg aacatgttaa gtcttttgaa
     3961 aacaaatact ctatgaggat ttatgagtgg ttattaaaag aactaacaca aaagaaaact
     4021 cacaaggcaa atatagagat tagccttgat gaatttaagt tcatgttaat gcttgaaaat
     4081 aactaccatg agtttaaaag gcttaaccaa tgggttttga aaccaataag taaagatttg
     4141 aacacttaca gcaatatgaa attggtggtt gataagcgag gccgcccgac tgatacgttg
     4201 attttccaag ttgaactaga tagacaaatg gatctcgtaa ccgaacttga gaacaaccag
     4261 ataaaaatga atggtgacaa aataccaaca accattacat cagattccta cctacataac
     4321 ggactaagaa aaacactaca cgatgcttta actgcaaaaa ttcagttcac cagttttgag
     4381 gcaaaatttt tgggtgacat gcaaagtaag tatgatctca atggttcgtt ctcatggctc
     4441 acgcaaaaac aacgaaccac actagagaac atactggcta aatacggaag gatctgaggt
     4501 tcttatggct cttgtatcta tcagtgaagc atcaagacta acaaacaaaa gtagaacaac
     4561 tgttcaccgt tacatatcaa agggaaaact gtccatatgc acagatgaaa acggtgtaaa
     4621 aaagatagat acatcagagc ttttacgagt ttttggtgca ttcaaagctg ttcaccatga
     4681 acagatcgac aatgtaacta ctagagctaa atacattcaa atatctatcc gctcatgaga
     4741 caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag tattcaacat
     4801 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4861 gaaacgctgg tgaaagtaaa agatgccgaa gatcagttgg gcgcacgtgt gggttacatc
     4921 gaactggacc tcaacagcgg taagattctt gagagttttc gccccgaaga acgtttccca
     4981 atgatgagca cttttaaagt tctgctctgt ggcgcggtat tatcccgtat tgacgccggg
     5041 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5101 gtcacagaaa agcatcttac ggacggcatg acagtacgcg aattatgcag cgctgccata
     5161 accacgagtg ataacacggc ggccaactta cttctgacaa cgatcggggg accgaaggag
     5221 cttaccgctt ttttgcacaa catgggtgat catgtaactc gccttgatcg ttgggaaccg
     5281 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agctatggca
     5341 acaacgttgc gcaaactctt aactggcgaa cttcttactc tcgcttcccg gcaacaatta
     5401 atagactgga tggaggcgga taaagttgcg ggaccacttc tgcgctcggc ccttccggct
     5461 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcccgcgg tattattgca
     5521 gccctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag
     5581 gcaactatgg acgaacgtaa tcgccagatc gctgagatag gtgcctccct gattaagcat
     5641 tggtaataat actagctccg gcaaaaaaac gggcaa
//
