LOCUS       sequence-324297-        3053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive superfolder GFP (sfGFP) expression, controlled under 
            E.coli J23106 (medium) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3053)
  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 3053)
  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: DT038A
FEATURES             Location/Qualifiers
     source          1..3053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     275..294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(513..617)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     685..703
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(707..750)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    753..772
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        774..808
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             837..860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             861..1574
                     /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..3053,1..443,444..512))
                     /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 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
       61 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
      121 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
      181 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
      241 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
      301 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
      361 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
      421 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
      481 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
      541 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
      601 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
      661 attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa
      721 aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agatttacgg
      781 ctagctcagt cctaggtata gtgctagcga tactagaaga aggagaataa tctatggact
      841 acaaagacga tgatgacaag atgcgtaaag gcgaagagct gttcactggt gtcgtcccta
      901 ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg
      961 aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc
     1021 cggtaccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt
     1081 atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc
     1141 aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat
     1201 ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg
     1261 gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg
     1321 ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg
     1381 gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc
     1441 tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga
     1501 aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg
     1561 atgaactgta caaataataa cgactcaggc tgctacgcct gtgtactgga aaacaaaacc
     1621 aaaacccaaa aaacaaaaaa ctgagcccat tggtatcgtg gaaggactcg atcaaaaaaa
     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 ggt
//
