LOCUS       sequence-389814-        3141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTFP1 under the control of PINA7 synthetic promoter, ColE1
            origin of replication, Ampicillin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3141)
  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 3141)
  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: pPINA7A2mTFP1(F)
FEATURES             Location/Qualifiers
     source          1..3141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..97
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    120..136
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    143..159
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             247..258
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             265..975
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     terminator      987..1073
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1346..1934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1426..1445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2022..2116)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2146..2937,2938..3006))
                     /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     2769..2788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3007..3111)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctactgggac gaagacgaac agtcgacaac gaagacgcca acatcaagat ctttattatc
       61 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca cctcgagaat
      121 tgtgagcgga taacaattga cattgtgagc ggataacaag atactgagca catctacctc
      181 tggcggtgat agatgattac ctctggcggt gatagatgat tacctctggc ggtgatagaa
      241 ttcattaaag aggagaaagg taccatggtg agcaagggcg aggagaccac aatgggcgta
      301 atcaagcccg acatgaagat caagctgaag atggagggca acgtgaatgg ccacgccttc
      361 gtgatcgagg gcgagggcga gggcaagccc tacgacggca ccaacaccat caacctggag
      421 gtgaaggagg gagcccccct gcccttctcc tacgacattc tgaccaccgc gttcgcctac
      481 ggcaacaggg ccttcaccaa gtaccccgac gacatcccca actacttcaa gcagtccttc
      541 cccgagggct actcttggga gcgcaccatg accttcgagg acaagggcat cgtgaaggtg
      601 aagtccgaca tctccatgga ggaggactcc ttcatctacg agatacacct caagggcgag
      661 aacttccccc ccaacggccc cgtgatgcag aagaagacca ccggctggga cgcctccacc
      721 gagaggatgt acgtgcgcga cggcgtgctg aagggcgacg tcaagcacaa gctgctgctg
      781 gagggcggcg gccaccaccg cgttgacttc aagaccatct acagggccaa gaaggcggtg
      841 aagctgcccg actatcactt tgtggaccac cgcatcgaga tcctgaacca cgacaaggac
      901 tacaacaagg tgaccgttta cgagagcgcc gtggcccgca actccaccga cggcatggac
      961 gagctgtaca agtaatctag aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1021 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc
     1081 cctagaccta ggcgatgacg gaaaaagaga tcgtcttaat gtatagattt tcgtggatta
     1141 cgtcgccagt caggatcctg gcagaagcta ttcattctgc agttccgtgc tccaaaacaa
     1201 ccgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1981 aaaactcacg ttaagggatt ttggtcatga ctagtgcttg gattctcacc aataaaaaac
     2041 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
     2101 tctatcaaca ggagtccaag cgagctcgta aacttggtct gacagttacc aatgcttaat
     2161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3121 ccgaaaagtg ccacctgacg t
//
