LOCUS       sequence-204381-        3716 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses tdTomato (a DsRed derivative) in Ecoli off a T7 promoter 
            when pNP1 tdTomato trigger is co-expressed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3716)
  AUTHORS   Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, 
            Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ
  TITLE     BioBits Explorer: A modular synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. 
            eCollection 2018 Aug.
  PUBMED    30083608
REFERENCE   2  (bases 1 to 3716)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5105. doi"; pages: " 
            10.1126/sciadv.aat5105. eCollection 2018 Aug"
COMMENT     Sequence Label: pNP1 tdTomato toehold switch
FEATURES             Location/Qualifiers
     source          1..3716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             126..1556
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(1694..1712)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1708..1755
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1973..2561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2053..2072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2733..3524,3525..3593))
                     /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     3356..3375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3594..3698)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggtagat attgaatgct
       61 gctgttatgt cgttaaacag aggagataac gaatgaacag cagcaacctg gcggcagcgc
      121 aaaagatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca
      181 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      241 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
      301 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
      361 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      421 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
      481 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
      541 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
      601 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
      661 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
      721 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
      781 agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc ggcagctccg
      841 gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg cgcttcaagg
      901 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      961 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     1021 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
     1081 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
     1141 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg
     1201 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
     1261 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
     1321 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccgc tacctggtgg
     1381 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
     1441 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     1501 gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagtaatgag
     1561 gatcccggga attctcgagt aaggttaacc tgcaggaggc ctttaattaa ggtggtgcgg
     1621 ccgcgctagc ggtcccgggg gatcgatccg gctgctaaca aagcccgaaa ggaagctgag
     1681 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1741 ttgaggggtt ttttgctgaa aggaggaact atatccggaa gcttggcact ggccgaccgg
     1801 ggtcgagcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1861 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2161 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2281 ttgagtccaa cccgctaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2401 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2581 ctacggggtc tgacgctcag tggaacgaaa actcacagat ccgggatttt ggtcatgaga
     2641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagt
//
