LOCUS       sequence-323504-        5917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ST6GAL1 with a GFP tag in its C-terminus in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5917)
  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 5917)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..5917
                     /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      1858..1867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1864..1866,1867..1869,1870..2583)
                     /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(1909..1930)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2705..2826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2742..2761)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2796..2815
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2833..3288)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2970..2991)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3182..3201
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3315..3419
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3421..3778
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3421..3612
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3444..3464)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3629..3764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3813..4607
                     /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             3813..4607
                     /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(3867..3886)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4477..4496
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4795..4814)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4839..4886
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5215..5803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5704..5723
                     /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 ttcccagtct gtatcctcaa gcagcaccca ggacccccac
      841 aggggccgcc agaccctcgg cagtctcaga ggcctagcca aggccaaacc agaggcctcc
      901 ttccaggtgt ggaacaagga cagctcttcc aaaaacctta tccctaggct gcaaaagatc
      961 tggaagaatt acctaagcat gaacaagtac aaagtgtcct acaaggggcc aggaccaggc
     1021 atcaagttca gtgcagaggc cctgcgctgc cacctccggg accatgtgaa tgtatccatg
     1081 gtagaggtca cagattttcc cttcaatacc tctgaatggg agggttatct gcccaaggag
     1141 agcattagga ccaaggctgg gccttggggc aggtgtgctg ttgtgtcgtc agcgggatct
     1201 ctgaagtcct cccaactagg cagagaaatc gatgatcatg acgcagtcct gaggtttaat
     1261 ggggcaccca cagccaactt ccaacaagat gtgggcacaa aaactaccat tcgcctgatg
     1321 aactctcagt tggttaccac agagaagcgc ttcctcaaag acagtttgta caatgaagga
     1381 atcctaattg tatgggaccc atctgtatac cactcagata tcccaaagtg gtaccagaat
     1441 ccggattata atttctttaa caactacaag acttatcgta agctgcaccc caatcagccc
     1501 ttttacatcc tcaagcccca gatgccttgg gagctatggg acattcttca agaaatctcc
     1561 ccagaagaga ttcagccaaa ccccccatcc tctgggatgc ttggtatcat catcatgatg
     1621 acgctgtgtg accaggtgga tatttatgag ttcctcccat ccaagcgcaa gactgacgtg
     1681 tgctactact accagaagtt cttcgatagt gcctgcacga tgggtgccta ccacccgctg
     1741 ctctatgaga agaatttggt gaagcatctc aaccagggca cagatgagga catctacctg
     1801 cttggaaaag ccacactgcc tggcttccgg accattcact gccgggatcc accggtcgcc
     1861 accatggtgt ccaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1921 gacggcgacg taaacggcca caagttctcc gtgcggggcg agggcgaggg cgatgccacc
     1981 aacggcaagc tgaccctgaa gttcatcagc accaccggca agctgcccgt gccctggccc
     2041 accctcgtga ccaccctgac ctacggcgtg cagagcttct cccgctaccc cgaccacatg
     2101 aagcgccacg acttcttcaa gagcgccatg cccgaaggct acgtccagga gcgcaccatc
     2161 tccttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2221 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2281 cacaagctgg agtacaactt caactcccac aacgtctata tcaccgccga caagcagaag
     2341 aacggcatca aggccaactt caagatccgc cacaacgtgg aggacggctc cgtgcagctc
     2401 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2461 cactacctgt ccacccagtc caagctgtcc aaagacccca acgagaagcg cgatcacatg
     2521 gtccttctgg aattcgtgac cgccgccggg atcactcacg gcatggacga gctgtacaag
     2581 taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2641 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2701 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2761 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2821 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2881 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2941 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3001 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3061 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3121 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3181 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3241 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3301 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3361 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3421 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3541 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3601 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3781 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3841 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3901 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3961 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4021 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4081 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4141 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4201 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4261 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4321 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4381 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4441 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4501 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4561 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4621 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4681 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4741 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4801 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4861 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4921 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4981 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5041 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5101 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5161 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5221 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5281 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5341 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5401 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5461 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5521 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5581 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5641 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5701 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5761 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5821 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5881 ccctgattct gtggataacc gtattaccgc catgcat
//
