LOCUS       sequence-177636-        5911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express SialT-AcGFP1 in mammalian cells, SialT encodes the full 
            length coding sequence of beta-galactosamide 
            alpha-2,6-sialyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5911)
  AUTHORS   Tie HC, Mahajan D, Chen B, Cheng L, VanDongen AM, Lu L
  TITLE     A novel imaging method for quantitative Golgi localization reveals 
            differential intra-Golgi trafficking of secretory cargos.
  JOURNAL   Mol Biol Cell. 2016 Jan 13. pii: mbc.E15-09-0664.
  PUBMED    26764092
REFERENCE   2  (bases 1 to 5911)
  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: "Mol Biol 
            Cell. 2016 Jan 13. pii: mbc.E15-09-0664."
COMMENT     Sequence Label: pAcGFP1-N1-SialT
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..134
                     /gene="bla"
                     /label=AmpR promoter
     promoter        136..493
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        136..327
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(159..179)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      344..479
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             528..1322
                     /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             528..1322
                     /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(582..601)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1192..1211
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1510..1529)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1554..1601
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1930..2518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2419..2438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2693..2996
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2997..3200
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3151..3171
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4484..4493
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4490..5209
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTMVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    5331..5452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5368..5387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5422..5441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(5459..5911,1..3))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5596..5617)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5808..5827
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
       61 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      121 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
      181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      241 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
      301 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
      361 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
      421 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
      481 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
      541 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
      601 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
      661 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
      721 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
      781 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
      841 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
      901 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
      961 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     1021 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     1081 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     1141 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     1201 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     1261 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     1321 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     1381 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     1441 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     1501 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     1561 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     1621 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     1681 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     1741 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     1801 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1861 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1921 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1981 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2041 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2101 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2161 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2221 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2281 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2341 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2401 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2461 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2521 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2581 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     2641 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     2701 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     2761 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2821 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     2881 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2941 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     3001 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     3061 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     3121 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     3181 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
     3241 agatctcgag atgattcaca ccaacctgaa gaaaaagttc agctgctgcg tcctggtctt
     3301 tcttctgttt gcagtcatct gtgtgtggaa ggaaaagaag aaagggagtt actatgattc
     3361 ctttaaattg caaaccaagg aattccaggt gttaaagagt ctggggaaat tggccatggg
     3421 gtctgattcc cagtctgtat cctcaagcag cacccaggac ccccacaggg gccgccagac
     3481 cctcggcagt ctcagaggcc tagccaaggc caaaccagag gcctccttcc aggtgtggaa
     3541 caaggacagc tcttccaaaa accttatccc taggctgcaa aagatctgga agaattacct
     3601 aagcatgaac aagtacaaag tgtcctacaa ggggccagga ccaggcatca agttcagtgc
     3661 agaggccctg cgctgccacc tccgggacca tgtgaatgta tccatggtag aggtcacaga
     3721 ttttcccttc aatacctctg aatgggaggg ttatctgccc aaggagagca ttaggaccaa
     3781 ggctgggcct tggggcaggt gtgctgttgt gtcgtcagcg ggatctctga agtcctccca
     3841 actaggcaga gaaatcgatg atcatgacgc agtcctgagg tttaatgggg cacccacagc
     3901 caacttccaa caagatgtgg gcacaaaaac taccattcgc ctgatgaact ctcagttggt
     3961 taccacagag aagcgcttcc tcaaagacag tttgtacaat gaaggaatcc taattgtatg
     4021 ggacccatct gtataccact cagatatccc aaagtggtac cagaatccgg attataattt
     4081 ctttaacaac tacaagactt atcgtaagct gcaccccaat cagccctttt acatcctcaa
     4141 gccccagatg ccttgggagc tatgggacat tcttcaagaa atctccccag aagagattca
     4201 gccaaacccc ccatcctctg ggatgcttgg tatcatcatc atgatgacgc tgtgtgacca
     4261 ggtggatatt tatgagttcc tcccatccaa gcgcaagact gacgtgtgct actactacca
     4321 gaagttcttc gatagtgcct gcacgatggg tgcctaccac ccgctgctct atgagaagaa
     4381 tttggtgaag catctcaacc agggcacaga tgaggacatc tacctgcttg gaaaagccac
     4441 actgcctggc ttccggacca ttcactgccg ggatccaccg gtcgccacca tggtgagcaa
     4501 gggcgccgag ctgttcaccg gcatcgtgcc catcctgatc gagctgaatg gcgatgtgaa
     4561 tggccacaag ttcagcgtga gcggcgaggg cgagggcgat gccacctacg gcaagctgac
     4621 cctgaagttc atctgcacca ccggcaagct gcctgtgccc tggcccaccc tggtgaccac
     4681 cctgagctac ggcgtgcagt gcttctcacg ctaccccgat cacatgaagc agcacgactt
     4741 cttcaagagc gccatgcctg agggctacat ccaggagcgc accatcttct tcgaggatga
     4801 cggcaactac aagtcgcgcg ccgaggtgaa gttcgagggc gataccatgg tgaatcgcat
     4861 cgagctgacc ggcaccgatt tcaaggagga tggcaacatc ctgggcaata agatggagta
     4921 caactacaac gcccacaatg tgtacatcat gaccgacaag gccaagaatg gcatcaaggt
     4981 gaacttcaag atccgccaca acatcgagga tggcagcgtg cagctggccg accactacca
     5041 gcagaatacc cccatcggcg atggccctgt gctgctgccc gataaccact acctgtccac
     5101 ccagagcgcc ctgtccaagg accccaacga gaagcgcgat cacatgatct acttcggctt
     5161 cgtgaccgcc gccgccatca cccacggcat ggatgagctg tacaagtaag cggccgcgac
     5221 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     5281 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     5341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     5461 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5521 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5581 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5641 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5701 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5761 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5821 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5881 cccgccgcgc ttaatgcgcc gctacagggc g
//
