LOCUS       sequence-158681-        5684 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Zipped GFP11 with TEV cleavage sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5684)
  AUTHORS   To TL, Schepis A, Ruiz-Gonzalez R, Zhang Q, Yu D, Dong Z, Coughlin 
            SR, Shu X
  TITLE     Rational Design of a GFP-Based Fluorogenic Caspase Reporter for 
            Imaging Apoptosis In Vivo.
  JOURNAL   Cell Chem Biol. 2016 Jul 21;23(7):875-82. doi: 
            10.1016/j.chembiol.2016.06.007.
  PUBMED    27447051
REFERENCE   2  (bases 1 to 5684)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chembiol.2016.06"; journalName: "Cell Chem Biol"; date: 
            "2016-07-21- 21"; volume: "23"; issue: "7"; pages: "875-82"
COMMENT     Sequence Label: ZipGFP11_TEV
FEATURES             Location/Qualifiers
     source          1..5684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1055..1102
                     /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"
     CDS             1103..1123
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1278..1295)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1284..1508
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1554..1982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1641..1660)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1851..1872
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1991..2011)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1996..2325
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2176..2311
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2238..2257
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2392..3186
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2446..2465)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3056..3075
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3360..3481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3397..3416)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3451..3470
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3530..3546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3530..3546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3543..3565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3554..3570
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3578..3584,3585..3602,3603..3608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3623..3644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3761..3778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3932..4517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4012..4031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4688..5479,5480..5548))
                     /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"
     primer_bind     5311..5330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5549..5653)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgggcgg cagcaaggtg tctgcactta aagaaaaggt
      961 ttccgcactc aaggagaaag tttcagcttt gaaagagaag gtcagtgccc ttaaggaaaa
     1021 agtgagcgcg ctgaaagaag gaggcagcgg atcccgggac cacatggtgc tgcacgagta
     1081 cgtgaacgcc gccggcatca cagaaaacct gtactttcag tccctcgagg gaggcagcga
     1141 agtgagcgcg ctggaaaaag aggtctccgc tcttgagaag gaagtttctg ccttggagaa
     1201 agaggtgagc gccctggaga aggaagtctc tgcgcttgaa aaataatcta gagggcccgt
     1261 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1321 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1381 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1441 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     1501 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     1561 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1621 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1681 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1741 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1801 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1861 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1921 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1981 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2041 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2101 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2161 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2221 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     2281 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     2341 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     2401 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2461 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2521 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     2581 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2641 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2701 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     2761 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     2821 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     2881 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     2941 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3001 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3061 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3121 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3181 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3241 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3301 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3361 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3421 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3481 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3541 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3601 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3661 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3721 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3781 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3841 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3901 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3961 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4021 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4081 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4141 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4201 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4261 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4321 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4381 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4441 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     4501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4561 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4621 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4681 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4741 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4801 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4861 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4921 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4981 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5041 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5101 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5161 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5221 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5281 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5341 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5401 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5461 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5521 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5581 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5641 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
