LOCUS       sequence-199339-        2486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of sfGFP from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2486)
  AUTHORS   Stark JC, Huang A, Nguyen PQ, Dubner RS, Hsu KJ, Ferrante TC, 
            Anderson M, Kanapskyte A, Mucha Q, Packett JS, Patel P, Patel R, Qaq
            D, Zondor T, Burke J, Martinez T, Miller-Berry A, Puppala A, 
            Reichert K, Schmid M, Brand L, Hill LR, Chellaswamy JF, Faheem N, 
            Fetherling S, Gong E, Gonzalzles EM, Granito T, Koritsaris J, Nguyen
            B, Ottman S, Palffy C, Patel A, Skweres S, Slaton A, Woods T, 
            Donghia N, Pardee K, Collins JJ, Jewett MC
  TITLE     BioBits Bright: A fluorescent synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5107. doi: 10.1126/sciadv.aat5107. 
            eCollection 2018 Aug.
  PUBMED    30083609
REFERENCE   2  (bases 1 to 2486)
  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: "2018-08-1"; volume: "4(8):eaat5107. doi"; pages: " 
            10.1126/sciadv.aat5107. eCollection 2018 Aug"
COMMENT     Sequence Label: pJL1-sfGFP
FEATURES             Location/Qualifiers
     source          1..2486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(166..185)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(167..185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(213..801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(293..312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(912..931)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             923..1738
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(996..1015)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(1756..1803)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1799..1817
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1872..1895)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(1896..1943)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
ORIGIN
        1 gccttcacct tcaccacgca cgctgaattt gtgaccgttc acatcgccat ccagttccac
       61 cagaatcggc acaacgccgg taaacagttc ttcacctttg ctcatatgta tatctccttc
      121 ttaaagttaa acaaaattat ttctagaggg aaaccgttgt ggtctcccta tagtgagtcg
      181 tattaatttc gcgggatcgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      241 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      301 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      361 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      421 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      481 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      541 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      601 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      661 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      721 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      781 cgcagaaaaa aaggatctca agaagatcct ttgatctcac gttgtgtctc aaaatctctg
      841 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
      901 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgaagcc
      961 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1021 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     1081 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1141 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1201 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     1261 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     1321 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     1381 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     1441 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
     1501 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1561 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1621 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1681 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1741 agaattggct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     1801 tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc
     1861 cggtcgactt atttttcgaa ctgcggatgg ctccacgtaa tacctgccgc attcacatat
     1921 tcgtgcagaa ccatgtggtc ccgcgtgcct ttttcgttcg gatctttaga cagaacggtc
     1981 tgcgtgctca gataatgatt atccggcagc agcaccggac catcaccaat cggggtattc
     2041 tgctgataat gatctgccag ctgcacactg ccatcttcaa cgttatggcg aacggtaaaa
     2101 ttcgctttga tgccattttt ctgtttatcc gccgtaatat aaacattatg gctattaaag
     2161 ttgtattcca gtttatggcc caggatattg ccatcttctt taaaatccgt gcctttcagt
     2221 tcaatgcggt tcaccagggt atcgccttca aatttcacaa cggcgcgcgt tttatatttg
     2281 ccatcatctt taaagctaat cgtacgttcc tgcacatagc cttccggcat tgcagattta
     2341 aagaaatcgt gacgtttcat gtgatccgga tagcgactaa aacactgaac gccataggtc
     2401 agggtggtca ccagcgtcgg ccacggcacc ggcagtttgc cggtggtgca gataaatttc
     2461 agcgtcagtt tgccaatcgt ggcatc
//
