LOCUS       sequence-322673-        6034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged MGAT1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6034)
  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 6034)
  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: MGAT1-GFP
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1991..2000
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1997..1999,2000..2002,2003..2716)
                     /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(2042..2063)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2838..2959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2875..2894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2929..2948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2966..3421)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3103..3124)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3315..3334
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3448..3552
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3554..3911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3554..3745
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3577..3597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3762..3897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3946..4740
                     /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             3946..4740
                     /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(4000..4019)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4610..4629
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4928..4947)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4972..5019
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5348..5936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5837..5856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gcgctaccgg actcagatct cgaggccacc atgctgaaga agcagtctgc
      661 agggcttgtg ctgtggggcg ctatcctctt tgtggcctgg aatgccctgc tgctcctctt
      721 cttctggacg cgcccagcac ctggcaggcc accctcagtc agcgctctcg atggcgaccc
      781 cgccagcctc acccgggaag tgattcgcct ggcccaagac gccgaggtgg agctggagcg
      841 gcagcgtggg ctgctgcagc agatcgggga tgccctgtcg agccagcggg ggagggtgcc
      901 caccgcggcc cctcccgccc agccgcgtgt gcctgtgacc cccgcgccgg cggtgattcc
      961 catcctagtc atcgcctgtg accgcagcac tgttcggcgc tgcctggaca agctgctgca
     1021 ttatcggccc tcggctgagc tcttccccat catcgttagc caggactgcg ggcacgagga
     1081 gacggcccag gccatcgcct cctacggcag cgcggtcacg cacatccggc agcccgacct
     1141 gagcagcatt gcggtgccgc cggaccaccg caagttccag ggctactaca agatcgcgcg
     1201 ccactaccgc tgggcgctgg gccaggtctt ccggcagttt cgcttccccg cggccgtggt
     1261 ggtggaggat gacctggagg tggccccgga cttcttcgag tactttcggg ccacctatcc
     1321 gctgctgaag gccgacccct ccctgtggtg cgtctcggcc tggaatgaca acggcaagga
     1381 gcagatggtg gacgccagca ggcctgagct gctctaccgc accgactttt tccctggcct
     1441 gggctggctg ctgttggccg agctctgggc tgagctggag cccaagtggc caaaggcctt
     1501 ctgggacgac tggatgcggc ggccggagca gcggcagggg cgggcctgca tacgccctga
     1561 gatctcaaga acgatgacct ttggccgcaa gggtgtgagc cacgggcagt tctttgacca
     1621 gcacctcaag tttatcaagc tgaaccagca gtttgtgcac ttcacccagc tggacctgtc
     1681 ttacctgcag cgggaggcct atgaccgaga tttcctcgcc cgcgtctacg gtgctcccca
     1741 gctgcaggtg gagaaagtga ggaccaatga ccggaaggag ctgggggagg tgcgggtgca
     1801 gtatacgggc agggacagct tcaaggcttt cgccaaggct ctgggtgtca tggatgacct
     1861 taagtcgggg gttccgagag ctggctaccg gggtattgtc accttccagt tccggggccg
     1921 ccgtgtccac ctggcgcccc caccgacgtg ggagggctat gatcctagct ggaatcggga
     1981 tccaccggtc gccaccatgg tgtccaaggg cgaggagctg ttcaccgggg tggtgcccat
     2041 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc tccgtgcggg gcgagggcga
     2101 gggcgatgcc accaacggca agctgaccct gaagttcatc agcaccaccg gcaagctgcc
     2161 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagagct tctcccgcta
     2221 ccccgaccac atgaagcgcc acgacttctt caagagcgcc atgcccgaag gctacgtcca
     2281 ggagcgcacc atctccttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt
     2341 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2401 caacatcctg gggcacaagc tggagtacaa cttcaactcc cacaacgtct atatcaccgc
     2461 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaacg tggaggacgg
     2521 ctccgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2581 gctgcccgac aaccactacc tgtccaccca gtccaagctg tccaaagacc ccaacgagaa
     2641 gcgcgatcac atggtccttc tggaattcgt gaccgccgcc gggatcactc acggcatgga
     2701 cgagctgtac aagtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2761 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2821 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2881 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2941 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3001 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3061 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3121 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3181 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3241 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3301 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3361 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3421 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3481 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3541 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3661 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3781 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3841 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3901 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3961 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4021 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4081 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4141 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4201 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4261 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4321 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4381 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4441 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4501 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4561 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4621 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4681 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4741 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4801 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4861 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4921 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4981 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5041 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5101 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5161 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5221 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5281 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5341 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5401 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5461 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5521 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5581 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5641 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5701 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5761 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5821 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5881 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5941 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6001 ttcctgcgtt atcccctgat tctgtggata accg
//
