LOCUS       sequence-250351-        4389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FKBP-CFAST11 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4389)
  AUTHORS   Tebo AG, Gautier A
  TITLE     A split fluorescent reporter with rapid and reversible 
            complementation.
  JOURNAL   Nat Commun. 2019 Jun 27;10(1):2822. doi: 10.1038/s41467-019-10855-0.
  PUBMED    31249300
REFERENCE   2  (bases 1 to 4389)
  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 
            Commun."; date: "2019-06-27"; volume: "10(1):2822. doi"; pages: " 
            10.1038/s41467-019-10855-0"
COMMENT     Sequence Label: pAG153 FKBP-CFAST11
FEATURES             Location/Qualifiers
     source          1..4389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             624..653
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             660..980
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    1177..1298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1214..1233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1268..1287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1305..1760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1442..1463)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1654..1673
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1787..1891
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1893..2250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1893..2084
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1916..1936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2101..2236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2285..3079
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2285..3079
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2339..2358)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2949..2968
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3267..3286)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3311..3358
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3687..4275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4176..4195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctgccacc atggaacaaa agcttatttc tgaagaggac ttggaattcg
      661 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
      721 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaatttgat tcctcccggg
      781 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
      841 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
      901 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
      961 tggagcttct aaaactggaa gaatccggag gaggcggcag cggcggaggg ggatccggtg
     1021 acagctattg ggtctttgtg aaacgggtgt aataggcggc cgcgactcta gatcataatc
     1081 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1141 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1261 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     1321 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     1381 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     1441 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     1501 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     1561 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     1621 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     1681 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     1741 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1861 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     1921 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1981 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2041 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2101 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2161 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2221 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     2281 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2341 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2401 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2461 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     2521 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     2581 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     2641 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     2701 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     2761 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     2821 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     2881 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     2941 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3001 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3061 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3121 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3181 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3241 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3301 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     3361 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     3421 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     3481 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     3541 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3601 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3661 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3721 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3781 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3841 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3901 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3961 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4021 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4081 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4141 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4201 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4261 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4321 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4381 gccatgcat
//
