LOCUS       sequence-324317-        3071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive superfolder GFP (sfGFP) expression, controlled under 
            E.coli tet promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3071)
  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 3071)
  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: DT036A
FEATURES             Location/Qualifiers
     source          1..3071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     257..276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(495..599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     667..685
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(689..732)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    735..754
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     protein_bind    756..774
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    781..799
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             838..861
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             862..1575
                     /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      1700..1747
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1749..1769
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1775..1792)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1946..2534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2026..2045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2705..3071,1..425,426..494))
                     /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 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
       61 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      121 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      181 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      241 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      301 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      361 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      421 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      481 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      541 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      601 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
      661 tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct
      721 aaggatgatt tctggaattc gcggccgctt ctagatccct atcagtgata gagattgaca
      781 tccctatcag tgatagagat actgagcacg atactagaag aaggagaata atctatggac
      841 tacaaagacg atgatgacaa gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct
      901 attctggtgg aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt
      961 gaaggtgacg caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg
     1021 ccggtacctt ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt
     1081 tatccggacc atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg
     1141 caggaacgca cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa
     1201 tttgaaggcg ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagac
     1261 ggcaatatcc tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc
     1321 gccgataaac aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat
     1381 ggcagcgtgc agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt
     1441 ctgctgccag acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag
     1501 aaacgcgatc atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg
     1561 gatgaactgt acaaataata acgactcagg ctgctacgcc tgtgtactgg aaaacaaaac
     1621 caaaacccaa aaaacaaaaa actgagccca ttggtatcgt ggaaggactc gatcaaaaaa
     1681 aaaaaaaaaa aaaaaaaaac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1741 ttttttggta ctagtagcgg ccgctgcagg cttcctcgct cactgactcg ctgcgctcgg
     1801 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1861 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1921 gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca
     1981 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2041 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2101 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2161 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2221 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2281 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2341 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2401 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2461 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2521 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2581 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2641 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2701 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2761 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2821 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2881 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2941 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3001 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3061 cattcagctc c
//
