LOCUS       sequence-324293-        3060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive superfolder GFP (sfGFP) expression, controlled under 
            E.coli hybrid J23117-J23106 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3060)
  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 3060)
  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: DT040A
FEATURES             Location/Qualifiers
     source          1..3060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     283..302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(521..625)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     693..711
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(715..758)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    761..780
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             845..868
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             869..1582
                     /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      1715..1762
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1764..1784
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1790..1807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1961..2549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2041..2060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2720..3060,1..451,452..520))
                     /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 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
       61 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
      121 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
      181 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
      241 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
      301 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
      361 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
      421 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
      481 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
      541 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
      601 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
      661 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca
      721 gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg ccgcttctag
      781 atttacggct agctcagtcc tagggattgt gctagcgata ctagaagaag gagaataatc
      841 tatggactac aaagacgatg atgacaagat gcgtaaaggc gaagagctgt tcactggtgt
      901 cgtccctatt ctggtggaac tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg
      961 cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg aagttcatct gtactactgg
     1021 taaactgccg gtaccttggc cgactctggt aacgacgctg acttatggtg ttcagtgctt
     1081 tgctcgttat ccggaccata tgaagcagca tgacttcttc aagtccgcca tgccggaagg
     1141 ctatgtgcag gaacgcacga tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga
     1201 agtgaaattt gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa
     1261 agaagacggc aatatcctgg gccataagct ggaatacaat tttaacagcc acaatgttta
     1321 catcaccgcc gataaacaaa aaaatggcat taaagcgaat tttaaaattc gccacaacgt
     1381 ggaggatggc agcgtgcagc tggctgatca ctaccagcaa aacactccaa tcggtgatgg
     1441 tcctgttctg ctgccagaca atcactatct gagcacgcaa agcgttctgt ctaaagatcc
     1501 gaacgagaaa cgcgatcata tggttctgct ggagttcgta accgcagcgg gcatcacgca
     1561 tggtatggat gaactgtaca aataataacg actcaggctg ctacgcctgt gtactggaaa
     1621 acaaaaccaa aacccaaaaa acaaaaaact gagcccattg gtatcgtgga aggactcgat
     1681 caaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaactagca taaccccttg gggcctctaa
     1741 acgggtcttg aggggttttt tggtactagt agcggccgct gcaggcttcc tcgctcactg
     1801 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1861 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1921 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc
     1981 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2161 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2281 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2341 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2401 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2581 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2641 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2701 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2881 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
//
