LOCUS       sequence-322820-        5965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Exprsses the GPI-anchored luminal domain of ST6GAL1 with a mCherry 
            tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5965)
  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 5965)
  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: ss-ED-mCherry-GPI
FEATURES             Location/Qualifiers
     source          1..5965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1829..2533
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1971..1989)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2239..2258
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2754..2875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2791..2810)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2845..2864
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2882..3337)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3019..3040)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3231..3250
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3364..3468
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3470..3827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3470..3661
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3493..3513)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3678..3813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3862..4656
                     /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             3862..4656
                     /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(3916..3935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4526..4545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4844..4863)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4888..4935
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5264..5852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5753..5772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      601 accggactca gatctcgagg ccaccatgta caggatgcaa ctcctgtctt gcattgcact
      661 aagtcttgca cttgtcacga aggaaaagaa gaaagggagt tactatgatt cctttaaatt
      721 gcaaaccaag gaattccagg tgttaaagag tctggggaaa ttggccatgg ggtctgattc
      781 ccagtctgta tcctcaagca gcacccagga cccccacagg ggccgccaga ccctcggcag
      841 tctcagaggc ctagccaagg ccaaaccaga ggcctccttc caggtgtgga acaaggacag
      901 ctcttccaaa aaccttatcc ctaggctgca aaagatctgg aagaattacc taagcatgaa
      961 caagtacaaa gtgtcctaca aggggccagg accaggcatc aagttcagtg cagaggccct
     1021 gcgctgccac ctccgggacc atgtgaatgt atccatggta gaggtcacag attttccctt
     1081 caatacctct gaatgggagg gttatctgcc caaggagagc attaggacca aggctgggcc
     1141 ttggggcagg tgtgctgttg tgtcgtcagc gggatctctg aagtcctccc aactaggcag
     1201 agaaatcgat gatcatgacg cagtcctgag gtttaatggg gcacccacag ccaacttcca
     1261 acaagatgtg ggcacaaaaa ctaccattcg cctgatgaac tctcagttgg ttaccacaga
     1321 gaagcgcttc ctcaaagaca gtttgtacaa tgaaggaatc ctaattgtat gggacccatc
     1381 tgtataccac tcagatatcc caaagtggta ccagaatccg gattataatt tctttaacaa
     1441 ctacaagact tatcgtaagc tgcaccccaa tcagcccttt tacatcctca agccccagat
     1501 gccttgggag ctatgggaca ttcttcaaga aatctcccca gaagagattc agccaaaccc
     1561 cccatcctct gggatgcttg gtatcatcat catgatgacg ctgtgtgacc aggtggatat
     1621 ttatgagttc ctcccatcca agcgcaagac tgacgtgtgc tactactacc agaagttctt
     1681 cgatagtgcc tgcacgatgg gtgcctacca cccgctgctc tatgagaaga atttggtgaa
     1741 gcatctcaac cagggcacag atgaggacat ctacctgctt ggaaaagcca cactgcctgg
     1801 cttccggacc attcactgcc gggatccggt gagcaagggc gaggaggata acatggccat
     1861 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     1921 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     1981 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     2041 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     2101 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     2161 gacccaggac tcctccctac aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     2221 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     2281 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     2341 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     2401 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     2461 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     2521 cgagctgtac aagggccggc cactggaaaa tggcggaacc tctctgtctg aaaaaactgt
     2581 gctgctgctg gtgactccct ttctggccgc tgcttggtcc ctccacccat gagcggccgc
     2641 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2701 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2761 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2821 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     2881 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2941 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3001 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3061 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3121 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3181 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3241 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3301 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3361 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3421 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3481 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4801 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4861 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4921 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4981 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5041 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5101 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5161 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5221 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5281 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5341 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5401 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5461 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5521 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5581 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5641 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5701 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5761 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5821 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5881 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5941 tggataaccg tattaccgcc atgca
//
