LOCUS       sequence-389741-        2931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E2-Crimson under the control of PANI2 synthetic promoter, 
            ColE1 origin of replication, Ampicillin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2931)
  AUTHORS   Sarkar K, Bonnerjee D, Srivastava R, Bagh S
  TITLE     A single layer artificial neural network type architecture with 
            molecular engineered bacteria for reversible and irreversible 
            computing.
  JOURNAL   Chem Sci. 2021 Nov 9;12(48):15821-15832. doi: 10.1039/d1sc01505b. 
            eCollection 2021 Dec 15.
  PUBMED    35024106
REFERENCE   2  (bases 1 to 2931)
  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: 
            "10.1039/d1sc01505b"; journalName: "Chem Sci"; date: "2021-11-9- 9";
            volume: "12"; issue: "48"; pages: "15821-15832"
COMMENT     Sequence Label: pPANI2A2E2-Crimson
FEATURES             Location/Qualifiers
     source          1..2931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    50..68
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    80..98
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             108..119
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             126..803
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     524..543
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      815..901
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1066..1654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1146..1165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1742..1836)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1866..2657,2658..2726))
                     /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     2489..2508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2727..2831)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2899..2917
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtacc tctggcggtg atagattacc tctggcggtg atattgacat ccctatcagt
       61 gatagagata ctgagcacat ccctatcagt gatagagaga attcattaaa gaggagaaag
      121 gtaccatgga tagcactgag aacgtcatca agcccttcat gcgcttcaag gtgcacatgg
      181 agggctccgt gaacggccac gagttcgaga tcgagggcgt gggcgagggc aagccctacg
      241 agggcaccca gaccgccaag ctgcaagtga ccaagggcgg ccccctgccc ttcgcctggg
      301 acatcctgtc cccccagttc ttctacggct ccaaggcgta catcaagcac cccgccgaca
      361 tccccgacta cctcaagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
      421 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcaccctca
      481 tctaccacgt gaagttcatc ggcgtgaact tcccctccga cggccccgta atgcagaaga
      541 agactctggg ctgggagccc tccactgagc gcaactaccc ccgcgacggc gtgctgaagg
      601 gcgagaacca catggcgctg aagctgaagg gcggcggcca ctacctgtgt gagttcaagt
      661 ccatctacat ggccaagaag cccgtgaagc tgcccggcta ccactacgtg gactacaagc
      721 tcgacatcac ctcccacaac gaggactaca ccgtggtgga gcagtacgag cgcgccgagg
      781 cccgccacca cctgttccag tagtctagag gcatcaaata aaacgaaagg ctcagtcgaa
      841 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
      901 tccgccgccc tagacctagg gcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1501 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga ctagtgcttg
     1741 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     1801 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgta aacttggtct
     1861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2461 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2821 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2881 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
