LOCUS       sequence-250352-        4386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FKBP-CFAST10 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4386)
  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 4386)
  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: pAG241 FKBP-CFAST10
FEATURES             Location/Qualifiers
     source          1..4386
                     /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    1174..1295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1211..1230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1265..1284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1302..1757)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1439..1460)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1651..1670
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1784..1888
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1890..2247
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1890..2081
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1913..1933)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2098..2233
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2282..3076
                     /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             2282..3076
                     /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(2336..2355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2946..2965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3264..3283)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3308..3355
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3684..4272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4173..4192
                     /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 aaacggtaat aggcggccgc gactctagat cataatcagc
     1081 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1141 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1201 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1261 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     1321 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1381 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1441 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1501 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1561 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1621 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1681 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1741 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1801 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1861 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     1921 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     1981 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2041 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2101 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2161 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2221 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     2281 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2341 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2401 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2461 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2521 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2581 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2641 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2701 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2761 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     2821 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     2881 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     2941 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3001 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3061 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3121 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3181 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3241 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3301 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     3361 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3421 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3481 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     3541 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3601 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3661 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3721 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3781 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3841 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3901 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3961 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4021 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4081 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4141 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4201 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4261 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4321 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4381 atgcat
//
