LOCUS       sequence-326571-        5521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive LuxR expression, controlled under T7 bacteriophage 
            promoter, and inducible PFO-sfGFP chimeric protein 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 5521)
  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 5521)
  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: DT047A
FEATURES             Location/Qualifiers
     source          1..5521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..54
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(58..101)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    104..123
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     primer_bind     125..144
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        125..143
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             174..197
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1072..1119
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1212..1235
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1236..1949
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3519..3566
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3568..3588
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(3594..3611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3765..4353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3845..3864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4524..5315,5316..5384))
                     /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     5147..5166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5385..5489)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt
       61 tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg cggccgcttc
      121 tagataatac gactcactat agggagatac tagaagaagg agaataatct atggactaca
      181 aagacgatga tgacaagatg aaaaacataa atgccgacga cacatacaga ataattaata
      241 aaattaaagc ttgtagaagc aataatgata ttaatcaatg cttatctgat atgactaaaa
      301 tggtacattg tgaatattat ttactcgcga tcatttatcc tcattctatg gttaaatctg
      361 atatttcaat cctagataat taccctaaaa aatggaggca atattatgat gacgctaatt
      421 taataaaata tgatcctata gtagattatt ctaactccaa tcattcacca attaattgga
      481 atatatttga aaacaatgct gtaaataaaa aatctccaaa tgtaattaaa gaagcgaaaa
      541 catcaggtct tatcactggg tttagtttcc ctattcatac ggctaacaat ggcttcggaa
      601 tgcttagttt tgcacattca gaaaaagaca actatataga tagtttattt ttacatgcgt
      661 gtatgaacat accattaatt gttccttctc tagttgataa ttatcgaaaa ataaatatag
      721 caaataataa atcaaacaac gatttaacca aaagagaaaa agaatgttta gcgtgggcat
      781 gcgaaggaaa aagctcttgg gatatttcaa aaatattagg ttgcagtgag cgtactgtca
      841 ctttccattt aaccaatgcg caaatgaaac tcaatacaac aaaccgctgc caaagtattt
      901 ctaaagcaat tttaacagga gcaattgatt gcccatactt taaaaattaa taacgactca
      961 ggctgctacg cctgtgtact ggaaaacaaa accaaaaccc aaaaaacaaa aaactgagcc
     1021 cattggtatc gtggaaggac tcgatcaaaa aaaaaaaaaa aaaaaaaaaa actagcataa
     1081 ccccttgggg cctctaaacg ggtcttgagg ggttttttgg tactagaacc tgtaggatcg
     1141 tacaggttta cgcaagaaaa tggtttgtta tagtcgaata aaaaatacta gaagaaggag
     1201 aataatctat ggactacaaa gacgatgatg acaagatgcg taaaggcgaa gagctgttca
     1261 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     1321 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     1381 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     1441 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     1501 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     1561 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     1621 actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     1681 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     1741 acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg
     1801 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     1861 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     1921 tcacgcatgg tatggatgaa ctgtacaaag gcggtggctc gggcggtggc tcgatgaaag
     1981 atatcacgga taaaaaccaa tcgattgata gtggcatcag ttcgctgagc tataaccgca
     2041 atgaagtcct ggccagcaat ggtgataaaa ttgaatcgtt tgttcctaaa gagggtaaaa
     2101 aagcgggcaa taaattcatt gtagtcgaac gccagaaacg tagcctgact acttcaccgg
     2161 ttgatatttc cattattgac agtgttaatg atcgcacgta tcctggtgcg ctgcaactgg
     2221 ctgataaagc atttgtcgaa aatcgcccta ctattctgat ggtaaaacgt aaaccgatca
     2281 atatcaacat tgatctgccg ggtctgaaag gtgaaaactc tatcaaggtg gatgatccaa
     2341 cttatggtaa agtcagcggc gccattgatg aactggttag caaatggaat gaaaaatata
     2401 gcagcaccca caccctgccg gcgcgcaccc agtactccga atccatggtg tatagtaaat
     2461 cccaaatctc atcggccctg aatgtgaacg ctaaagtgct ggaaaattcg ctgggtgttg
     2521 acttcaacgc cgtagcaaat aacgagaaaa aagtaatgat cctggcatat aagcagatct
     2581 tttatacagt cagtgccgat ctgccgaaaa atccgtctga tctgttcgat gatagcgtta
     2641 cttttaatga tctgaagcaa aaaggtgtta gcaacgaagc tccgccgctg atggtttcta
     2701 atgttgccta tggtcgcacc atttatgtta aactggaaac cactagctcc tcgaaagatg
     2761 tccaagctgc ttttaaagct ctgatcaaaa atacagatat taaaaatagt caacagtata
     2821 aagatattta tgaaaatagc agtttcacag cagttgtgct gggaggggat gcacaggaac
     2881 ataataaagt ggtgacgaaa gattttgacg aaatccgcaa agtcatcaaa gataatgcca
     2941 cttttagtac gaagaatccg gcttatccaa tcagctatac ctctgtcttc ctgaaagaca
     3001 acagcgttgc cgccgtccat aacaagaccg attacattga aaccaccagt accgaatatt
     3061 ctaaaggtaa aatcaacctg gatcattccg gtgcgtatgt cgctcagttt gaagtggcgt
     3121 gggacgaggt ctcatacgat aaagaaggta acgaagttct gacgcataaa acttgggacg
     3181 gcaattacca ggacaaaact gcccactata gcaccgttat cccgctggaa gccaatgcgc
     3241 gcaacatccg cattaaagca cgtgaagcaa cggggctggc ctgggaatgg tggcgtgatg
     3301 taattagcga atatgatgtg ccgctgacga ataatatcaa cgtttcgatt tgggggacca
     3361 ccctgtatcc gggttcaagc attacctata attgatgacg actcaggctg ctacgcctgt
     3421 gtactggaaa acaaaaccaa aacccaaaaa acaaaaaact gagcccattg gtatcgtgga
     3481 aggactcgat caaaaaaaaa aaaaaaaaaa aaaaaaaact agcataaccc cttggggcct
     3541 ctaaacgggt cttgaggggt tttttggtac tagtagcggc cgctgcaggc ttcctcgctc
     3601 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3661 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3721 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca caggctccgc
     3781 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3841 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3901 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3961 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4021 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4081 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4141 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4201 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4261 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4321 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4381 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4441 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4501 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4561 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4621 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4681 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4741 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4801 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4861 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4921 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4981 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5041 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5101 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5161 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5221 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5281 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5341 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5401 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5461 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5521 t
//
