LOCUS       sequence-177295-        4233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express GalT-HA in mammalian cells, GalT encodes the 1-81 amino 
            acids from beta-1,4-galactosyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4233)
  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 4233)
  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: pHA-N1-GalT
FEATURES             Location/Qualifiers
     source          1..4233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..159
                     /gene="bla"
                     /label=AmpR promoter
     promoter        161..518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        161..352
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(184..204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      369..504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             553..1347
                     /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             553..1347
                     /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(607..626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1217..1236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1535..1554)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1579..1626
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1955..2543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2444..2463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2718..3021
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3022..3225
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3176..3196
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3530..3556
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3678..3799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3715..3734)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3769..3788
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(3806..4233,1..28))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3943..3964)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4155..4174
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
       61 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      121 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
      181 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
      241 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
      301 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
      361 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
      421 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
      481 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
      541 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
      601 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
      661 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
      721 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
      781 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
      841 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
      901 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
      961 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     1021 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     1081 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     1141 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     1201 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     1261 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     1321 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     1381 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     1441 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     1501 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     1561 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     1621 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     1681 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     1741 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     1801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     1861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2581 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2641 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     2701 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     2761 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     2821 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     2881 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     2941 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     3001 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     3061 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     3121 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     3181 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     3241 cagatccgct agcatgaggc ttcgggagcc gctcctgagc ggcagcgccg cgatgccagg
     3301 cgcgtcccta cagcgggcct gccgcctgct cgtggccgtc tgcgctctgc accttggcgt
     3361 caccctcgtt tactacctgg ctggccgcga cctgagccgc ctgccccaac tggtcggagt
     3421 ctccacaccg ctgcagggcg gctcgaacag tgccgccgcc atcgggcagt cctccgggga
     3481 gctccggacc ggaggggcca aggatccacc ggtcggatcc gccaccatgt acccatacga
     3541 tgttccagat tacgctgcgg ccgcgactct agatcataat cagccatacc acatttgtag
     3601 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3661 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3721 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3781 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3841 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3901 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3961 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4021 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4081 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4141 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4201 gcggtcacgc tgcgcgtaac caccacaccc gcc
//
