LOCUS       sequence-326569-        4812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive LuxR expression, controlled under E.coli tet promoter, 
            and inducible Perfringolysin O (PFO) expression, controlled under V.
            fischeri HSL-regulated pLux (luxPR) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4812)
  AUTHORS   Toparlak OD, Zasso J, Bridi S, Serra MD, Macchi P, Conti L, Baudet 
            ML, Mansy SS
  TITLE     Artificial cells drive neural differentiation.
  JOURNAL   Sci Adv. 2020 Sep 18;6(38). pii: 6/38/eabb4920. doi: 
            10.1126/sciadv.abb4920. Print 2020 Sep.
  PUBMED    32948587
REFERENCE   2  (bases 1 to 4812)
  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: "Sci Adv."; 
            date: "2020-09-18"; volume: "6(38). pii: 6/38/eabb4920. doi"; pages:
            " 10.1126/sciadv.abb4920. Print 2020 Sep"
COMMENT     Sequence Label: DT046A
FEATURES             Location/Qualifiers
     source          1..4812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..63
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(67..110)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    113..132
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     protein_bind    134..152
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    159..177
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             216..239
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1112..1159
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1252..1275
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2819..2866
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2868..2888
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(2894..2911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3065..3653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3145..3164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3824..4615,4616..4684))
                     /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     4447..4466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4685..4789)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
       61 ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc
      121 ggccgcttct agatccctat cagtgataga gattgacatc cctatcagtg atagagatac
      181 tgagcacgat actagaagaa ggagaataat ctatggacta caaagacgat gatgacaaga
      241 tgaaaaacat aaatgccgac gacacataca gaataattaa taaaattaaa gcttgtagaa
      301 gcaataatga tattaatcaa tgcttatctg atatgactaa aatggtacat tgtgaatatt
      361 atttactcgc gatcatttat cctcattcta tggttaaatc tgatatttca atcctagata
      421 attaccctaa aaaatggagg caatattatg atgacgctaa tttaataaaa tatgatccta
      481 tagtagatta ttctaactcc aatcattcac caattaattg gaatatattt gaaaacaatg
      541 ctgtaaataa aaaatctcca aatgtaatta aagaagcgaa aacatcaggt cttatcactg
      601 ggtttagttt ccctattcat acggctaaca atggcttcgg aatgcttagt tttgcacatt
      661 cagaaaaaga caactatata gatagtttat ttttacatgc gtgtatgaac ataccattaa
      721 ttgttccttc tctagttgat aattatcgaa aaataaatat agcaaataat aaatcaaaca
      781 acgatttaac caaaagagaa aaagaatgtt tagcgtgggc atgcgaagga aaaagctctt
      841 gggatatttc aaaaatatta ggttgcagtg agcgtactgt cactttccat ttaaccaatg
      901 cgcaaatgaa actcaataca acaaaccgct gccaaagtat ttctaaagca attttaacag
      961 gagcaattga ttgcccatac tttaaaaatt aataacgact caggctgcta cgcctgtgta
     1021 ctggaaaaca aaaccaaaac ccaaaaaaca aaaaactgag cccattggta tcgtggaagg
     1081 actcgatcaa aaaaaaaaaa aaaaaaaaaa actagcataa ccccttgggg cctctaaacg
     1141 ggtcttgagg ggttttttgg tactagaacc tgtaggatcg tacaggttta cgcaagaaaa
     1201 tggtttgtta tagtcgaata aaaaatacta gaagaaggag aataatctat ggactacaaa
     1261 gacgatgatg acaagatgaa agatatcacg gataaaaacc aatcgattga tagtggcatc
     1321 agttcgctga gctataaccg caatgaagtc ctggccagca atggtgataa aattgaatcg
     1381 tttgttccta aagagggtaa aaaagcgggc aataaattca ttgtagtcga acgccagaaa
     1441 cgtagcctga ctacttcacc ggttgatatt tccattattg acagtgttaa tgatcgcacg
     1501 tatcctggtg cgctgcaact ggctgataaa gcatttgtcg aaaatcgccc tactattctg
     1561 atggtaaaac gtaaaccgat caatatcaac attgatctgc cgggtctgaa aggtgaaaac
     1621 tctatcaagg tggatgatcc aacttatggt aaagtcagcg gcgccattga tgaactggtt
     1681 agcaaatgga atgaaaaata tagcagcacc cacaccctgc cggcgcgcac ccagtactcc
     1741 gaatccatgg tgtatagtaa atcccaaatc tcatcggccc tgaatgtgaa cgctaaagtg
     1801 ctggaaaatt cgctgggtgt tgacttcaac gccgtagcaa ataacgagaa aaaagtaatg
     1861 atcctggcat ataagcagat cttttataca gtcagtgccg atctgccgaa aaatccgtct
     1921 gatctgttcg atgatagcgt tacttttaat gatctgaagc aaaaaggtgt tagcaacgaa
     1981 gctccgccgc tgatggtttc taatgttgcc tatggtcgca ccatttatgt taaactggaa
     2041 accactagct cctcgaaaga tgtccaagct gcttttaaag ctctgatcaa aaatacagat
     2101 attaaaaata gtcaacagta taaagatatt tatgaaaata gcagtttcac agcagttgtg
     2161 ctgggagggg atgcacagga acataataaa gtggtgacga aagattttga cgaaatccgc
     2221 aaagtcatca aagataatgc cacttttagt acgaagaatc cggcttatcc aatcagctat
     2281 acctctgtct tcctgaaaga caacagcgtt gccgccgtcc ataacaagac cgattacatt
     2341 gaaaccacca gtaccgaata ttctaaaggt aaaatcaacc tggatcattc cggtgcgtat
     2401 gtcgctcagt ttgaagtggc gtgggacgag gtctcatacg ataaagaagg taacgaagtt
     2461 ctgacgcata aaacttggga cggcaattac caggacaaaa ctgcccacta tagcaccgtt
     2521 atcccgctgg aagccaatgc gcgcaacatc cgcattaaag cacgtgaagc aacggggctg
     2581 gcctgggaat ggtggcgtga tgtaattagc gaatatgatg tgccgctgac gaataatatc
     2641 aacgtttcga tttgggggac caccctgtat ccgggttcaa gcattaccta taattgatga
     2701 cgactcaggc tgctacgcct gtgtactgga aaacaaaacc aaaacccaaa aaacaaaaaa
     2761 ctgagcccat tggtatcgtg gaaggactcg atcaaaaaaa aaaaaaaaaa aaaaaaaact
     2821 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttggtac tagtagcggc
     2881 cgctgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3061 cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4801 gaaaagtgcc ac
//
