LOCUS       sequence-121003-        4938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF7-ZF8 Op 1gap -- GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4938)
  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 4938)
  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-163
FEATURES             Location/Qualifiers
     source          1..4938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        66..97
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      124..133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(130..132,133..135,136..849)
                     /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(130..132,133..135,136..846)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(175..196)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(436..455)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     783..804
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    893..1014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(904..923)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     958..977
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1208..1565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1208..1399
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1231..1251)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1416..1551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1478..1497
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1596..2633
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2612..2620
                     /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    2657..2705
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2861..2878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3032..3620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3112..3131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3820..4611,4612..4680))
                     /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    4785..4833
                     /note="synthetic polyadenylation signal"
     misc_feature    4847..4938
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4887..4906
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcagctggag ggtgaagtgg tccaagctta
       61 gacactagag ggtatataat ggaagctcga cttccagctt ggcaatccgg tactgttggt
      121 aaagccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      181 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      241 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      301 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
      361 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      421 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      481 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      541 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
      601 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
      661 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      721 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
      781 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
      841 tacaagtaat aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
      901 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
      961 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1021 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1081 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc
     1141 ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     1201 ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     1261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1381 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     1441 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1501 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1561 gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta
     1621 ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt
     1681 cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc
     1741 gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca
     1801 gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat
     1861 actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc
     1921 ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc
     1981 aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg
     2041 atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca
     2101 ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc
     2161 ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg
     2221 gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg
     2281 tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact
     2341 tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca
     2401 tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg
     2461 ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa
     2521 tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg
     2581 gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag
     2641 aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     2701 gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2761 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2821 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     2881 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2941 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3001 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3061 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3121 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3181 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3241 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3301 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3361 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3421 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3481 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3541 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3601 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3661 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3721 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3781 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
     3841 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
     3901 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
     3961 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
     4021 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
     4081 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
     4141 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
     4201 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
     4261 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
     4321 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
     4381 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
     4441 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
     4501 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
     4561 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
     4621 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
     4681 actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg
     4741 ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa atatctttat
     4801 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
     4861 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     4921 gtgccagaac atttctct
//
