LOCUS       sequence-322815-        5345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FKBP tagged partial length ST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5345)
  AUTHORS   Sun X, Mahajan D, Chen B, Song Z, Lu L
  TITLE     A quantitative study of the Golgi retention of glycosyltransferases.
  JOURNAL   J Cell Sci. 2021 Oct 15;134(20). pii: 272560. doi: 
            10.1242/jcs.258564. Epub 2021 Oct 21.
  PUBMED    34533190
REFERENCE   2  (bases 1 to 5345)
  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: "J Cell 
            Sci."; date: "2021-10-15"; volume: "134(20). pii: 272560. doi"; 
            pages: " 10.1242/jcs.258564. Epub 2021 Oct 21"
COMMENT     Sequence Label: FKBP-ST*-GFP
FEATURES             Location/Qualifiers
     source          1..5345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(607..609,610..927)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     regulatory      1288..1297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1294..1296,1297..1299,1300..2013)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1339..1360)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2135..2256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2172..2191)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2226..2245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2263..2718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2400..2421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2612..2631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2745..2849
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2851..3208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2851..3042
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2874..2894)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3059..3194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3243..4037
                     /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             3243..4037
                     /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(3297..3316)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3907..3926
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4225..4244)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4269..4316
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4645..5233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5134..5153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gccaccatgg tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc
      661 ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa atttgattcc
      721 tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc
      781 tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca
      841 gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc
      901 ttcgatgtgg agcttctaaa actggaactc gaggccacca tgattcacac caacctgaag
      961 aaaaagttca gctgctgcgt actagtcttt cttctgtttg cagtcatatg tgtgtggaag
     1021 gaaaagaaga aagggagtta ctatgattcc tttaaattgc aaaccaagga attccaggtg
     1081 ttaaagagtc tggggaaatt ggccatgggg tctgattccc agtctgtatc ctcaagcagc
     1141 acccaggacc cccacagggg ccgccagacc ctcggcagtc tcagaggcct agccaaggcc
     1201 aaaccagagg cctccttcca ggtgtggaac aaggacagct cttccaaaaa ccttatccct
     1261 aggctgcaaa agatctgggc accggtcgcc accatggtgt ccaagggcga ggagctgttc
     1321 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttctcc
     1381 gtgcggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatcagc
     1441 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1501 cagagcttct cccgctaccc cgaccacatg aagcgccacg acttcttcaa gagcgccatg
     1561 cccgaaggct acgtccagga gcgcaccatc tccttcaagg acgacggcac ctacaagacc
     1621 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1681 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt caactcccac
     1741 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1801 cacaacgtgg aggacggctc cgtgcagctc gccgaccact accagcagaa cacccccatc
     1861 ggcgacggcc ccgtgctgct gcccgacaac cactacctgt ccacccagtc caagctgtcc
     1921 aaagacccca acgagaagcg cgatcacatg gtccttctgg aattcgtgac cgccgccggg
     1981 atcactcacg gcatggacga gctgtacaag taagcggccg cgactctaga tcataatcag
     2041 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2101 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2161 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2221 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2281 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2341 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2401 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2461 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2521 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2581 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2641 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2701 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2761 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2821 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     2881 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2941 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3001 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3061 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3121 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3181 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3241 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3301 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3361 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3421 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3481 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3541 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3601 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3661 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3721 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     3781 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3841 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3901 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3961 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4021 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4081 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4141 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4201 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4261 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4321 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4381 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4441 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5341 catgc
//
