LOCUS       sequence-322784-        6823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with the cytosolic tail 
            from ST6GAL1, and TMD and luminal domain from TFR in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  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 6823)
  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: SFF-GFP
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(1445..1456)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2801..2810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2807..2809,2810..2812,2813..3526)
                     /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(2852..2873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3648..3769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3685..3704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3739..3758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3776..4231)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3913..3934)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4125..4144
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4258..4362
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4364..4721
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4364..4555
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4387..4407)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4572..4707
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4756..5550
                     /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             4756..5550
                     /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(4810..4829)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5420..5439
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5738..5757)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5782..5829
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6158..6746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6647..6666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgaggccac
      661 catgattcac accaacctga agaaaaagtg tagtggaagt atctgctatg ggactattgc
      721 tgtgatcgtc tttttcttga ttggatttat gattggctac ttgggctatt gtaaaggggt
      781 agaaccaaaa actgagtgtg agagactggc aggaaccgag tctccagtga gggaggagcc
      841 aggagaggac ttccctgcag cacgtcgctt atattgggat gacctgaaga gaaagttgtc
      901 ggagaaactg gacagcacag acttcaccag caccatcaag ctgctgaatg aaaattcata
      961 tgtccctcgt gaggctggat ctcaaaaaga tgaaaatctt gcgttgtatg ttgaaaatca
     1021 atttcgtgaa tttaaactca gcaaagtctg gcgtgatcaa cattttgtta agattcaggt
     1081 caaagacagc gctcaaaact cggtgatcat agttgataag aacggtagac ttgtttacct
     1141 ggtggagaat cctgggggtt atgtggcgta tagtaaggct gcaacagtta ctggtaaact
     1201 ggtccatgct aattttggta ctaaaaaaga ttttgaggat ttatacactc ctgtgaatgg
     1261 atctatagtg attgtcagag cagggaaaat cacctttgca gaaaaggttg caaatgctga
     1321 aagcttaaat gcaattggtg tgttgatata catggaccag actaaatttc ccattgttaa
     1381 cgcagaactt tcattctttg gacatgctca tctggggaca ggtgaccctt acacacctgg
     1441 attcccttcc ttcaatcaca ctcagtttcc accatctcgg tcatcaggat tgcctaatat
     1501 acctgtccag acaatctcca gagctgctgc agaaaagctg tttgggaata tggaaggaga
     1561 ctgtccctct gactggaaaa cagactctac atgtaggatg gtaacctcag aaagcaagaa
     1621 tgtgaagctc actgtgagca atgtgctgaa agagataaaa attcttaaca tctttggagt
     1681 tattaaaggc tttgtagaac cagatcacta tgttgtagtt ggggcccaga gagatgcatg
     1741 gggccctgga gctgcaaaat ccggtgtagg cacagctctc ctattgaaac ttgcccagat
     1801 gttctcagat atggtcttaa aagatgggtt tcagcccagc agaagcatta tctttgccag
     1861 ttggagtgct ggagactttg gatcggttgg tgccactgaa tggctagagg gatacctttc
     1921 gtccctgcat ttaaaggctt tcacttatat taatctggat aaagcggttc ttggtaccag
     1981 caacttcaag gtttctgcca gcccactgtt gtatacgctt attgagaaaa caatgcaaaa
     2041 tgtgaagcat ccggttactg ggcaatttct atatcaggac agcaactggg ccagcaaagt
     2101 tgagaaactc actttagaca atgctgcttt ccctttcctt gcatattctg gaatcccagc
     2161 agtttctttc tgtttttgcg aggacacaga ttatccttat ttgggtacca ccatggacac
     2221 ctataaggaa ctgattgaga ggattcctga gttgaacaaa gtggcacgag cagctgcaga
     2281 ggtcgctggt cagttcgtga ttaaactaac ccatgatgtt gaattgaacc tggactatga
     2341 gaggtacaac agccaactgc tttcatttgt gagggatctg aaccaataca gagcagacat
     2401 aaaggaaatg ggcctgagtt tacagtggct gtattctgct cgtggagact tcttccgtgc
     2461 tacttccaga ctaacaacag atttcgggaa tgctgagaaa acagacagat ttgtcatgaa
     2521 gaaactcaat gatcgtgtca tgagagtgga gtatcacttc ctctctccct acgtatctcc
     2581 aaaagagtct cctttccgac atgtcttctg gggctccggc tctcacacgc tgccagcttt
     2641 actggagaac ttgaaactgc gtaaacaaaa taacggtgct tttaatgaaa cgctgttcag
     2701 aaaccagttg gctctagcta cttggactat tcagggagct gcaaatgccc tctctggtga
     2761 cgtttgggac attgacaatg agtttcggga tccaccggtc gccaccatgg tgtccaaggg
     2821 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2881 ccacaagttc tccgtgcggg gcgagggcga gggcgatgcc accaacggca agctgaccct
     2941 gaagttcatc agcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3001 gacctacggc gtgcagagct tctcccgcta ccccgaccac atgaagcgcc acgacttctt
     3061 caagagcgcc atgcccgaag gctacgtcca ggagcgcacc atctccttca aggacgacgg
     3121 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3181 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3241 cttcaactcc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     3301 cttcaagatc cgccacaacg tggaggacgg ctccgtgcag ctcgccgacc actaccagca
     3361 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgtccaccca
     3421 gtccaagctg tccaaagacc ccaacgagaa gcgcgatcac atggtccttc tggaattcgt
     3481 gaccgccgcc gggatcactc acggcatgga cgagctgtac aagtaagcgg ccgcgactct
     3541 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3601 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3781 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3841 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3901 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3961 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4021 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4081 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4141 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4201 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4261 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4321 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     4381 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4441 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4561 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4621 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4681 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4741 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4801 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4861 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4921 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4981 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5041 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5101 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5161 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5221 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5281 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5341 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5401 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5461 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5521 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5581 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5641 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5701 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5761 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5821 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5881 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5941 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     6001 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6121 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6181 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6241 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6301 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6361 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6421 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6481 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6541 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6601 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6661 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6721 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6781 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atc
//
