LOCUS       sequence-323507-        6004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a ST6GAL1 mutant with a GFP tag in its C-terminus in 
            mammalian cells. The mutated stem region of Tac is inserted within 
            ST..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6004)
  AUTHORS   Sun X, Tie HC, Chen B, Lu L
  TITLE     Glycans function as a Golgi export signal to promote the 
            constitutive exocytic trafficking.
  JOURNAL   J Biol Chem. 2020 Aug 21. pii: RA120.014476. doi: 
            10.1074/jbc.RA120.014476.
  PUBMED    32826314
REFERENCE   2  (bases 1 to 6004)
  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 Biol 
            Chem. 2020 Aug 21. pii: RA120.014476. doi: 
            10.1074/jbc.RA120.014476."
COMMENT     Sequence Label: ST-Tstem(5A)-GFP
FEATURES             Location/Qualifiers
     source          1..6004
                     /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      1945..1954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1951..1953,1954..1956,1957..2670)
                     /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(1996..2017)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2792..2913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2829..2848)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2883..2902
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2920..3375)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3057..3078)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3269..3288
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3402..3506
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3508..3865
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3508..3699
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3531..3551)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3716..3851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3900..4694
                     /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             3900..4694
                     /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(3954..3973)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4564..4583
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4882..4901)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4926..4973
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5302..5890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5791..5810
                     /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 atctcgaggc caccatgatt cacaccaacc tgaagaaaaa gttcagctgc
      661 tgcgtcctgg tctttcttct gtttgcagtc atctgtgtgt ggaaggaaaa gaagaaaggg
      721 agttactatg attcctttaa attgcaaacc aaggaattcc aggtgttaaa gagtctgggg
      781 aaattggcca tggggtctga ttccccgcgg ctcgtcacaa cagcagattt tcaaatacag
      841 gcagaaatgg ctgcagccat ggagacgtcc atatttgcag cagagtacca gggatcccag
      901 tctgtatcct caagcagcac ccaggacccc cacaggggcc gccagaccct cggcagtctc
      961 agaggcctag ccaaggccaa accagaggcc tccttccagg tgtggaacaa ggacagctct
     1021 tccaaaaacc ttatccctag gctgcaaaag atctggaaga attacctaag catgaacaag
     1081 tacaaagtgt cctacaaggg gccaggacca ggcatcaagt tcagtgcaga ggccctgcgc
     1141 tgccacctcc gggaccatgt gaatgtatcc atggtagagg tcacagattt tcccttcaat
     1201 acctctgaat gggagggtta tctgcccaag gagagcatta ggaccaaggc tgggccttgg
     1261 ggcaggtgtg ctgttgtgtc gtcagcggga tctctgaagt cctcccaact aggcagagaa
     1321 atcgatgatc atgacgcagt cctgaggttt aatggggcac ccacagccaa cttccaacaa
     1381 gatgtgggca caaaaactac cattcgcctg atgaactctc agttggttac cacagagaag
     1441 cgcttcctca aagacagttt gtacaatgaa ggaatcctaa ttgtatggga cccatctgta
     1501 taccactcag atatcccaaa gtggtaccag aatccggatt ataatttctt taacaactac
     1561 aagacttatc gtaagctgca ccccaatcag cccttttaca tcctcaagcc ccagatgcct
     1621 tgggagctat gggacattct tcaagaaatc tccccagaag agattcagcc aaacccccca
     1681 tcctctggga tgcttggtat catcatcatg atgacgctgt gtgaccaggt ggatatttat
     1741 gagttcctcc catccaagcg caagactgac gtgtgctact actaccagaa gttcttcgat
     1801 agtgcctgca cgatgggtgc ctaccacccg ctgctctatg agaagaattt ggtgaagcat
     1861 ctcaaccagg gcacagatga ggacatctac ctgcttggaa aagccacact gcctggcttc
     1921 cggaccattc actgcgcacc ggtcgccacc atggtgtcca agggcgagga gctgttcacc
     1981 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttctccgtg
     2041 cggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catcagcacc
     2101 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2161 agcttctccc gctaccccga ccacatgaag cgccacgact tcttcaagag cgccatgccc
     2221 gaaggctacg tccaggagcg caccatctcc ttcaaggacg acggcaccta caagacccgc
     2281 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2341 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa ctcccacaac
     2401 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     2461 aacgtggagg acggctccgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2521 gacggccccg tgctgctgcc cgacaaccac tacctgtcca cccagtccaa gctgtccaaa
     2581 gaccccaacg agaagcgcga tcacatggtc cttctggaat tcgtgaccgc cgccgggatc
     2641 actcacggca tggacgagct gtacaagtaa gcggccgcga ctctagatca taatcagcca
     2701 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2761 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2821 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2881 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     2941 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3001 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3061 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3121 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3181 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3241 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3301 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3361 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3421 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3481 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3541 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3601 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3661 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3721 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3781 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3841 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3901 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3961 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4021 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4081 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4141 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4201 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4261 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4321 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4381 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4441 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4501 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4561 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4621 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4681 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4741 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4801 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4861 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4921 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     4981 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5041 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5101 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5161 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5221 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5281 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5341 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5401 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5461 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5521 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5581 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5641 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5701 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5761 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5821 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5881 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5941 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6001 gcat
//
