LOCUS       sequence-324285-        3054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive superfolder GFP (sfGFP) expression, controlled under 
            E.coli tac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3054)
  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 3054)
  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: DT035A
FEATURES             Location/Qualifiers
     source          1..3054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     177..194
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    200..220
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(222..269)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(396..1109)
                     /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"
     CDS             complement(1110..1133)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(join(1167..1173,1174..1189,1190..1195))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_feature    1196..1217
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1220..1263
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1267..1285)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1353..1457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1458..1526,1527..2318)
                     /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     complement(1676..1695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     2978..2997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caggggggcg gagcctgtgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
       61 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      121 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      181 agtcagtgag cgaggaagcc tgcagcggcc gctactagta ccaaaaaacc cctcaagacc
      241 cgtttagagg ccccaagggg ttatgctagt tttttttttt tttttttttt ttttttgatc
      301 gagtccttcc acgataccaa tgggctcagt tttttgtttt ttgggttttg gttttgtttt
      361 ccagtacaca ggcgtagcag cctgagtcgt tattatttgt acagttcatc cataccatgc
      421 gtgatgcccg ctgcggttac gaactccagc agaaccatat gatcgcgttt ctcgttcgga
      481 tctttagaca gaacgctttg cgtgctcaga tagtgattgt ctggcagcag aacaggacca
      541 tcaccgattg gagtgttttg ctggtagtga tcagccagct gcacgctgcc atcctccacg
      601 ttgtggcgaa ttttaaaatt cgctttaatg ccattttttt gtttatcggc ggtgatgtaa
      661 acattgtggc tgttaaaatt gtattccagc ttatggccca ggatattgcc gtcttcttta
      721 aagtcaatgc ctttcagctc aatgcggttt accagggtat cgccttcaaa tttcacttcc
      781 gcacgcgttt tgtacgtgcc gtcatcctta aaggaaatcg tgcgttcctg cacatagcct
      841 tccggcatgg cggacttgaa gaagtcatgc tgcttcatat ggtccggata acgagcaaag
      901 cactgaacac cataagtcag cgtcgttacc agagtcggcc aaggtaccgg cagtttacca
      961 gtagtacaga tgaacttcag cgtcagttta ccattagttg cgtcaccttc accctcgcca
     1021 cgcacggaaa acttatgacc gttgacatca ccatccagtt ccaccagaat agggacgaca
     1081 ccagtgaaca gctcttcgcc tttacgcatc ttgtcatcat cgtctttgta gtccatagat
     1141 tattctcctt cttctagtat cccacacatt atacgagccg atgattaatt gtcaactcta
     1201 gaagcggccg cgaattccag aaatcatcct tagcgaaagc taaggatttt ttttatctga
     1261 aattctgcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1321 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1441 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1501 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1561 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1621 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1681 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1741 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1801 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1861 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1921 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1981 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2041 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2101 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2161 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2221 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2281 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2341 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2401 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2461 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2521 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2581 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2641 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2701 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2761 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2821 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2881 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2941 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3001 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgt
//
