LOCUS       sequence-324298-        3052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive superfolder GFP (sfGFP) expression, controlled under 
            E.coli J23101 (strong) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3052)
  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 3052)
  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: DT037A
FEATURES             Location/Qualifiers
     source          1..3052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     300..319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(538..642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     710..728
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(732..775)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    778..797
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        799..833
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     CDS             862..885
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             886..1599
                     /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      1724..1771
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1773..1793
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1799..1816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1970..2558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2050..2069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2729..3052,1..468,469..537))
                     /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"
ORIGIN
        1 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
       61 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
      121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
      181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
      241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
      301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
      361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
      421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
      481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
      541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
      601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
      661 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
      721 cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctggaa
      781 ttcgcggccg cttctagatt tacagctagc tcagtcctag gtattatgct agcgatacta
      841 gaagaaggag aataatctat ggactacaaa gacgatgatg acaagatgcg taaaggcgaa
      901 gagctgttca ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat
      961 aagttttccg tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag
     1021 ttcatctgta ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact
     1081 tatggtgttc agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag
     1141 tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg
     1201 tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg
     1261 aaaggcattg actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt
     1321 aacagccaca atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt
     1381 aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac
     1441 actccaatcg gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc
     1501 gttctgtcta aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc
     1561 gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat aataacgact caggctgcta
     1621 cgcctgtgta ctggaaaaca aaaccaaaac ccaaaaaaca aaaaactgag cccattggta
     1681 tcgtggaagg actcgatcaa aaaaaaaaaa aaaaaaaaaa aaactagcat aaccccttgg
     1741 ggcctctaaa cgggtcttga ggggtttttt ggtactagta gcggccgctg caggcttcct
     1801 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1861 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccacaggc
     1981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2521 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2581 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2641 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2701 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2881 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc at
//
