LOCUS       sequence-121006-        4970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF9 Op x4 -- GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4970)
  AUTHORS   Slomovic S, Collins JJ
  TITLE     DNA sense-and-respond protein modules for mammalian cells.
  JOURNAL   Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.
  PUBMED    26389572
REFERENCE   2  (bases 1 to 4970)
  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: "Nat 
            Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585."
COMMENT     Sequence Label: pGL4.26-SS-191
FEATURES             Location/Qualifiers
     source          1..4970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        98..129
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      156..165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(162..164,165..167,168..881)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(162..164,165..167,168..878)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(207..228)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(468..487)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     815..836
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    925..1046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(936..955)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     990..1009
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1240..1597
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1240..1431
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1263..1283)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1448..1583
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1510..1529
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1628..2665
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2644..2652
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    2689..2737
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2893..2910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3064..3652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3144..3163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3852..4643,4644..4712))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4817..4865
                     /note="synthetic polyadenylation signal"
     misc_feature    4879..4970
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4919..4938
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gctgtagatg gagaaatgta gatggagaaa
       61 tgtagatgga gaaatgtaga tggagaagct tagacactag agggtatata atggaagctc
      121 gacttccagc ttggcaatcc ggtactgttg gtaaagccac catggtgagc aagggcgagg
      181 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      241 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      301 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      361 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      421 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      481 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      541 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      601 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
      661 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
      721 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
      781 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
      841 ccgccgggat cactctcggc atggacgagc tgtacaagta ataattctag agtcggggcg
      901 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
      961 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1021 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1081 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     1141 tcgataagga tccgtttgcg tattgggcgc tcttccgctg atctgcgcag caccatggcc
     1201 tgaaataacc tctgaaagag gaacttggtt agctaccttc tgaggcggaa agaaccagct
     1261 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1321 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     1381 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1441 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1501 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     1561 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcgattctt ctgacactag
     1621 cgccaccatg aagaagcccg aactcaccgc taccagcgtt gaaaaatttc tcatcgagaa
     1681 gttcgacagt gtgagcgacc tgatgcagtt gtcggagggc gaagagagcc gagccttcag
     1741 cttcgatgtc ggcggacgcg gctatgtact gcgggtgaat agctgcgctg atggcttcta
     1801 caaagaccgc tacgtgtacc gccacttcgc cagcgctgca ctacccatcc ccgaagtgtt
     1861 ggacatcggc gagttcagcg agagcctgac atactgcatc agtagacgcg cccaaggcgt
     1921 tactctccaa gacctccccg aaacagagct gcctgctgtg ttacagcctg tcgccgaagc
     1981 tatggatgct attgccgccg ccgacctcag tcaaaccagc ggcttcggcc cattcgggcc
     2041 ccaaggcatc ggccagtaca caacctggcg ggatttcatt tgcgccattg ctgatcccca
     2101 tgtctaccac tggcagaccg tgatggacga caccgtgtcc gccagcgtag ctcaagccct
     2161 ggacgaactg atgctgtggg ccgaagactg tcccgaggtg cgccacctcg tccatgccga
     2221 cttcggcagc aacaacgtcc tgaccgacaa cggccgcatc accgccgtaa tcgactggtc
     2281 cgaagctatg ttcggggaca gtcagtacga ggtggccaac atcttcttct ggcggccctg
     2341 gctggcttgc atggagcagc agactcgcta cttcgagcgc cggcatcccg agctggccgg
     2401 cagccctcgt ctgcgagcct acatgctgcg catcggcctg gatcagctct accagagcct
     2461 cgtggacggc aacttcgacg atgctgcctg ggctcaaggc cgctgcgatg ccatcgtccg
     2521 cagcggggcc ggcaccgtcg gtcgcacaca aatcgctcgc cggagcgcag ccgtatggac
     2581 cgacggctgc gtcgaggtgc tggccgacag cggcaaccgc cggcccagta cacgaccgcg
     2641 cgctaaggag gtaggtcgag tttaaactct agaaccggtc atggccgcaa taaaatatct
     2701 ttattttcat tacatctgtg tgttggtttt ttgtgtgttc gaactagatg ctgtcgaccg
     2761 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     2821 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     2881 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2941 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3001 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3061 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3481 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agcggccgca aatgctaaac
     3841 cactgcagtg gttaccagtg cttgatcagt gaggcaccga tctcagcgat ctgcctattt
     3901 cgttcgtcca tagtggcctg actccccgtc gtgtagatca ctacgattcg tgagggctta
     3961 ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc gttcaccggc ccccgatttg
     4021 tcagcaatga accagccagc agggagggcc gagcgaagaa gtggtcctgc tactttgtcc
     4081 gcctccatcc agtctatgag ctgctgtcgt gatgctagag taagaagttc gccagtgagt
     4141 agtttccgaa gagttgtggc cattgctact ggcatcgtgg tatcacgctc gtcgttcggt
     4201 atggcttcgt tcaactctgg ttcccagcgg tcaagccggg tcacatgatc acccatatta
     4261 tgaagaaatg cagtcagctc cttagggcct ccgatcgttg tcagaagtaa gttggccgcg
     4321 gtgttgtcgc tcatggtaat ggcagcacta cacaattctc ttaccgtcat gccatccgta
     4381 agatgctttt ccgtgaccgg cgagtactca accaagtcgt tttgtgagta gtgtatacgg
     4441 cgaccaagct gctcttgccc ggcgtctata cgggacaaca ccgcgccaca tagcagtact
     4501 ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga aagactcaag gatcttgccg
     4561 ctattgagat ccagttcgat atagcccact cttgcaccca gttgatcttc agcatctttt
     4621 actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc aaagaaggga
     4681 atgagtgcga cacgaaaatg ttggatgctc atactcgtcc tttttcaata ttattgaagc
     4741 atttatcagg gttactagta cgtctctcaa ggataagtaa gtaatattaa ggtacgggag
     4801 gtattggaca ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4861 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     4921 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//
