LOCUS       sequence-322771-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with the cytosolic tail 
            from TFR and TMD and luminal domain from ST6GAL1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  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 6073)
  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: FSS-GFP
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2022..2031
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2028..2030,2031..2033,2034..2747)
                     /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(2073..2094)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2869..2990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2906..2925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2960..2979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2997..3452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3134..3155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3346..3365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3479..3583
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3585..3942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3585..3776
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3608..3628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3793..3928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3977..4771
                     /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             3977..4771
                     /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(4031..4050)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4641..4660
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4959..4978)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5003..5050
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5379..5967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5868..5887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgctacc ggactcagat ctcgaggcca ccatgatgga tcaagctaga tcagcattct
      661 ctaacttgtt tggtggagaa ccattgtcat atacccggtt cagcctggct cggcaagtag
      721 atggcgataa cagtcatgtg gagatgaaac ttgctgtaga tgaagaagaa aatgctgaca
      781 ataacacaaa ggccaatgtc acaaaaccaa aaaggttcag ctgctgcgtc ctggtctttc
      841 ttctgtttgc agtcatctgt gtgtggaagg aaaagaagaa agggagttac tatgattcct
      901 ttaaattgca aaccaaggaa ttccaggtgt taaagagtct ggggaaattg gccatggggt
      961 ctgattccca gtctgtatcc tcaagcagca cccaggaccc ccacaggggc cgccagaccc
     1021 tcggcagtct cagaggccta gccaaggcca aaccagaggc ctccttccag gtgtggaaca
     1081 aggacagctc ttccaaaaac cttatcccta ggctgcaaaa gatctggaag aattacctaa
     1141 gcatgaacaa gtacaaagtg tcctacaagg ggccaggacc aggcatcaag ttcagtgcag
     1201 aggccctgcg ctgccacctc cgggaccatg tgaatgtatc catggtagag gtcacagatt
     1261 ttcccttcaa tacctctgaa tgggagggtt atctgcccaa ggagagcatt aggaccaagg
     1321 ctgggccttg gggcaggtgt gctgttgtgt cgtcagcggg atctctgaag tcctcccaac
     1381 taggcagaga aatcgatgat catgacgcag tcctgaggtt taatggggca cccacagcca
     1441 acttccaaca agatgtgggc acaaaaacta ccattcgcct gatgaactct cagttggtta
     1501 ccacagagaa gcgcttcctc aaagacagtt tgtacaatga aggaatccta attgtatggg
     1561 acccatctgt ataccactca gatatcccaa agtggtacca gaatccggat tataatttct
     1621 ttaacaacta caagacttat cgtaagctgc accccaatca gcccttttac atcctcaagc
     1681 cccagatgcc ttgggagcta tgggacattc ttcaagaaat ctccccagaa gagattcagc
     1741 caaacccccc atcctctggg atgcttggta tcatcatcat gatgacgctg tgtgaccagg
     1801 tggatattta tgagttcctc ccatccaagc gcaagactga cgtgtgctac tactaccaga
     1861 agttcttcga tagtgcctgc acgatgggtg cctaccaccc gctgctctat gagaagaatt
     1921 tggtgaagca tctcaaccag ggcacagatg aggacatcta cctgcttgga aaagccacac
     1981 tgcctggctt ccggaccatt cactgccggg atccaccggt cgccaccatg gtgtccaagg
     2041 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2101 gccacaagtt ctccgtgcgg ggcgagggcg agggcgatgc caccaacggc aagctgaccc
     2161 tgaagttcat cagcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2221 tgacctacgg cgtgcagagc ttctcccgct accccgacca catgaagcgc cacgacttct
     2281 tcaagagcgc catgcccgaa ggctacgtcc aggagcgcac catctccttc aaggacgacg
     2341 gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2401 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2461 acttcaactc ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     2521 acttcaagat ccgccacaac gtggaggacg gctccgtgca gctcgccgac cactaccagc
     2581 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgtccaccc
     2641 agtccaagct gtccaaagac cccaacgaga agcgcgatca catggtcctt ctggaattcg
     2701 tgaccgccgc cgggatcact cacggcatgg acgagctgta caagtaagcg gccgcgactc
     2761 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2821 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3001 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3061 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3121 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3181 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3241 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3301 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3361 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3421 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3601 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3661 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3721 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3781 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3841 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3901 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3961 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4021 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4081 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4141 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4201 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4261 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4321 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4381 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4441 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4501 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4561 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4621 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4681 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4741 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4801 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4861 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4921 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4981 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5041 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5101 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5161 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5221 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5281 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5341 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5401 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5461 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5521 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5581 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5641 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5701 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5761 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5821 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5881 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5941 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6001 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6061 aaccgtatta ccg
//
