LOCUS       sequence-324320-        3043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive superfolder GFP (sfGFP) expression, controlled under T7
            bacteriophage promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3043)
  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 3043)
  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: DT043A
FEATURES             Location/Qualifiers
     source          1..3043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     273..292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(511..615)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     683..701
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(705..748)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    751..770
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     primer_bind     772..791
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        772..790
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             821..844
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             845..1558
                     /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      1688..1735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1737..1757
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1763..1780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1934..2522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2014..2033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2693..3043,1..441,442..510))
                     /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 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
       61 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
      481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
      541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
      661 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa
      721 atccttagct ttcgctaagg atgatttctg gaattcgcgg ccgcttctag ataatacgac
      781 tcactatagg gagatactag aagaaggaga ataatctatg gactacaaag acgatgatga
      841 caagatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga
      901 tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa
      961 tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac
     1021 tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa
     1081 gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc
     1141 ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct
     1201 ggtaaaccgc attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca
     1261 taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa
     1321 tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc
     1381 tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca
     1441 ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt
     1501 tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaata
     1561 ataacgactc aggctgctac gcctgtgtac tggaaaacaa aaccaaaacc caaaaaacaa
     1621 aaaactgagc ccattggtat cgtggaagga ctcgatcaaa aaaaaaaaaa aaaaaaaaaa
     1681 aaaaaaacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttggtact
     1741 agtagcggcc gctgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1801 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1861 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1921 cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2101 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2341 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tat
//
