LOCUS       sequence-317586-        6794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with a truncated 
            cytosolic tail from TFR, TMD from ST6GAL1 and luminal domain from 
            TFR in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6794)
  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 6794)
  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: F9SF-GFP
FEATURES             Location/Qualifiers
     source          1..6794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1378..1389)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2734..2743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2740..2742,2743..2745,2746..3459)
                     /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(2785..2806)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3582..3703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3619..3638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3673..3692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3710..4165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3847..3868)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4059..4078
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4192..4296
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4298..4655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4298..4489
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4321..4341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4506..4641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4690..5484
                     /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             4690..5484
                     /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(4744..4763)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5354..5373
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5672..5691)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5716..5763
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6092..6680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6581..6600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatggcc aatgtcacaa aaccaaaaag gttcagctgc
      661 tgcgtcctgg tctttcttct gtttgcagtc atctgtgtgt ggtgtaaagg ggtagaacca
      721 aaaactgagt gtgagagact ggcaggaacc gagtctccag tgagggagga gccaggagag
      781 gacttccctg cagcacgtcg cttatattgg gatgacctga agagaaagtt gtcggagaaa
      841 ctggacagca cagacttcac cagcaccatc aagctgctga atgaaaattc atatgtccct
      901 cgtgaggctg gatctcaaaa agatgaaaat cttgcgttgt atgttgaaaa tcaatttcgt
      961 gaatttaaac tcagcaaagt ctggcgtgat caacattttg ttaagattca ggtcaaagac
     1021 agcgctcaaa actcggtgat catagttgat aagaacggta gacttgttta cctggtggag
     1081 aatcctgggg gttatgtggc gtatagtaag gctgcaacag ttactggtaa actggtccat
     1141 gctaattttg gtactaaaaa agattttgag gatttataca ctcctgtgaa tggatctata
     1201 gtgattgtca gagcagggaa aatcaccttt gcagaaaagg ttgcaaatgc tgaaagctta
     1261 aatgcaattg gtgtgttgat atacatggac cagactaaat ttcccattgt taacgcagaa
     1321 ctttcattct ttggacatgc tcatctgggg acaggtgacc cttacacacc tggattccct
     1381 tccttcaatc acactcagtt tccaccatct cggtcatcag gattgcctaa tatacctgtc
     1441 cagacaatct ccagagctgc tgcagaaaag ctgtttggga atatggaagg agactgtccc
     1501 tctgactgga aaacagactc tacatgtagg atggtaacct cagaaagcaa gaatgtgaag
     1561 ctcactgtga gcaatgtgct gaaagagata aaaattctta acatctttgg agttattaaa
     1621 ggctttgtag aaccagatca ctatgttgta gttggggccc agagagatgc atggggccct
     1681 ggagctgcaa aatccggtgt aggcacagct ctcctattga aacttgccca gatgttctca
     1741 gatatggtct taaaagatgg gtttcagccc agcagaagca ttatctttgc cagttggagt
     1801 gctggagact ttggatcggt tggtgccact gaatggctag agggatacct ttcgtccctg
     1861 catttaaagg ctttcactta tattaatctg gataaagcgg ttcttggtac cagcaacttc
     1921 aaggtttctg ccagcccact gttgtatacg cttattgaga aaacaatgca aaatgtgaag
     1981 catccggtta ctgggcaatt tctatatcag gacagcaact gggccagcaa agttgagaaa
     2041 ctcactttag acaatgctgc tttccctttc cttgcatatt ctggaatccc agcagtttct
     2101 ttctgttttt gcgaggacac agattatcct tatttgggta ccaccatgga cacctataag
     2161 gaactgattg agaggattcc tgagttgaac aaagtggcac gagcagctgc agaggtcgct
     2221 ggtcagttcg tgattaaact aacccatgat gttgaattga acctggacta tgagaggtac
     2281 aacagccaac tgctttcatt tgtgagggat ctgaaccaat acagagcaga cataaaggaa
     2341 atgggcctga gtttacagtg gctgtattct gctcgtggag acttcttccg tgctacttcc
     2401 agactaacaa cagatttcgg gaatgctgag aaaacagaca gatttgtcat gaagaaactc
     2461 aatgatcgtg tcatgagagt ggagtatcac ttcctctctc cctacgtatc tccaaaagag
     2521 tctcctttcc gacatgtctt ctggggctcc ggctctcaca cgctgccagc tttactggag
     2581 aacttgaaac tgcgtaaaca aaataacggt gcttttaatg aaacgctgtt cagaaaccag
     2641 ttggctctag ctacttggac tattcaggga gctgcaaatg ccctctctgg tgacgtttgg
     2701 gacattgaca atgagtttcg ggatccaccg gtcgccacca tggtgtccaa gggcgaggag
     2761 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2821 ttctccgtgc ggggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
     2881 atcagcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2941 ggcgtgcaga gcttctcccg ctaccccgac cacatgaagc gccacgactt cttcaagagc
     3001 gccatgcccg aaggctacgt ccaggagcgc accatctcct tcaaggacga cggcacctac
     3061 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3121 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
     3181 tcccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     3241 atccgccaca acgtggagga cggctccgtg cagctcgccg accactacca gcagaacacc
     3301 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgtccac ccagtccaag
     3361 ctgtccaaag accccaacga gaagcgcgat cacatggtcc ttctggaatt cgtgaccgcc
     3421 gccgggatca ctcacggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     3481 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3541 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3601 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3661 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3721 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3781 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3841 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3901 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3961 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4021 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4081 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4141 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4201 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4261 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4321 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4381 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4441 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4501 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4561 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4621 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4681 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4741 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4801 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4861 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4921 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4981 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5041 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5101 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5161 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5221 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5281 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5341 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5401 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5461 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5521 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5581 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5641 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5701 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5761 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5821 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5881 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5941 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6001 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6061 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6121 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6181 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     6241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6601 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6661 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6721 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6781 ttaccgccat gcat
//
