LOCUS       sequence-370632-        3704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tdTomato from PR promoter with optimized RBS (i2) in 
            E.coli Dh5alpha cell. The construct is in reverse direction network 
            brick..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3704)
  AUTHORS   Mukhopadhyay S, Sarkar K, Srivastava R, Pal A, Bagh S
  TITLE     Processing two environmental chemical signals with a synthetic 
            genetic IMPLY gate, a 2-input-2-output integrated logic circuit, and
            a process pipeline to optimize its systems chemistry in Escherichia 
            coli.
  JOURNAL   Biotechnol Bioeng. 2020 May;117(5):1502-1512. doi: 
            10.1002/bit.27286. Epub 2020 Feb 7.
  PUBMED    31981217
REFERENCE   2  (bases 1 to 3704)
  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: "10.1002/bit.27286";
            journalName: "Biotechnol Bioeng"; date: "2020-05"; volume: "117"; 
            issue: "5"; pages: "1502-1512"
COMMENT     Sequence Label: pNB_EA_R3tdTomatoi2
FEATURES             Location/Qualifiers
     source          1..3704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..137
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(138..206,207..998)
                     /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(356..375)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1028..1122
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1210..1798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1699..1718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      2080..2089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2086..3516
                     /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
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     terminator      3539..3625
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
       61 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      121 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      181 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      241 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      301 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      361 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
      421 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
      481 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
      541 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
      601 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
      661 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
      721 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
      781 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
      841 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
      901 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
      961 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttacgagc
     1021 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     1081 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcac tagtcatgac
     1141 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1201 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1261 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1321 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1381 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1441 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1501 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1561 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1621 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1681 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1741 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1801 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1861 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1921 taccgctcgc cgcagccgaa cggttgtttt ggagcacgga actgcagttc taccaggatc
     1981 ctggtaactc gagtaacacc gtgcgtgttg actattttac ctctggcggt gataatggtt
     2041 gcatgtacga attcacagca gacgagaaag gtaccggtcg ccaccatggt gagcaagggc
     2101 gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     2161 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2221 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     2281 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     2341 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     2401 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     2461 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2521 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     2581 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     2641 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     2701 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     2761 ctggggcatg gcaccggcag caccggcagc ggcagctccg gcaccgcctc ctccgaggac
     2821 aacaacatgg ccgtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg
     2881 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2941 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     3001 ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac
     3061 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3121 ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg
     3181 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     3241 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     3301 caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg
     3361 gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc
     3421 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     3481 ctgttcctgt acggcatgga cgagctgtac aagtaggcgg ccgcgactct agaggcatca
     3541 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3601 gaacgctctc ctgagtagga caaatccgcc gccctagacc taggaacaaa tgcgagatct
     3661 tgatgttggc gtcttcgttg tcgactgttc gtcttcgtcc cagt
//
