LOCUS       sequence-322818-        5947 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with a truncated 
            cytosolic tail from TFR, TMD from TFR and luminal domain from 
            ST6GAL1 in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5947)
  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 5947)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: F9FS-GFP
FEATURES             Location/Qualifiers
     source          1..5947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        78..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      636..645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1905..1914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1911..1913,1914..1916,1917..2630)
                     /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(1956..1977)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2752..2873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2789..2808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2843..2862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2880..3335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3017..3038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3229..3248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3362..3466
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3468..3825
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3468..3659
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3491..3511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3676..3811
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3860..4654
                     /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             3860..4654
                     /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(3914..3933)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4524..4543
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4842..4861)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4886..4933
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5262..5850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5751..5770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      601 cagatccgct agcgctaccg gactcagatc tcgaggccac catggccaat gtcacaaaac
      661 caaaaaggtg tagtggaagt atctgctatg ggactattgc tgtgatcgtc tttttcttga
      721 ttggatttat gattggctac ttgggctata aggaaaagaa gaaagggagt tactatgatt
      781 cctttaaatt gcaaaccaag gaattccagg tgttaaagag tctggggaaa ttggccatgg
      841 ggtctgattc ccagtctgta tcctcaagca gcacccagga cccccacagg ggccgccaga
      901 ccctcggcag tctcagaggc ctagccaagg ccaaaccaga ggcctccttc caggtgtgga
      961 acaaggacag ctcttccaaa aaccttatcc ctaggctgca aaagatctgg aagaattacc
     1021 taagcatgaa caagtacaaa gtgtcctaca aggggccagg accaggcatc aagttcagtg
     1081 cagaggccct gcgctgccac ctccgggacc atgtgaatgt atccatggta gaggtcacag
     1141 attttccctt caatacctct gaatgggagg gttatctgcc caaggagagc attaggacca
     1201 aggctgggcc ttggggcagg tgtgctgttg tgtcgtcagc gggatctctg aagtcctccc
     1261 aactaggcag agaaatcgat gatcatgacg cagtcctgag gtttaatggg gcacccacag
     1321 ccaacttcca acaagatgtg ggcacaaaaa ctaccattcg cctgatgaac tctcagttgg
     1381 ttaccacaga gaagcgcttc ctcaaagaca gtttgtacaa tgaaggaatc ctaattgtat
     1441 gggacccatc tgtataccac tcagatatcc caaagtggta ccagaatccg gattataatt
     1501 tctttaacaa ctacaagact tatcgtaagc tgcaccccaa tcagcccttt tacatcctca
     1561 agccccagat gccttgggag ctatgggaca ttcttcaaga aatctcccca gaagagattc
     1621 agccaaaccc cccatcctct gggatgcttg gtatcatcat catgatgacg ctgtgtgacc
     1681 aggtggatat ttatgagttc ctcccatcca agcgcaagac tgacgtgtgc tactactacc
     1741 agaagttctt cgatagtgcc tgcacgatgg gtgcctacca cccgctgctc tatgagaaga
     1801 atttggtgaa gcatctcaac cagggcacag atgaggacat ctacctgctt ggaaaagcca
     1861 cactgcctgg cttccggacc attcactgcc gggatccacc ggtcgccacc atggtgtcca
     1921 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1981 acggccacaa gttctccgtg cggggcgagg gcgagggcga tgccaccaac ggcaagctga
     2041 ccctgaagtt catcagcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2101 ccctgaccta cggcgtgcag agcttctccc gctaccccga ccacatgaag cgccacgact
     2161 tcttcaagag cgccatgccc gaaggctacg tccaggagcg caccatctcc ttcaaggacg
     2221 acggcaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2281 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2341 acaacttcaa ctcccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     2401 ccaacttcaa gatccgccac aacgtggagg acggctccgt gcagctcgcc gaccactacc
     2461 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgtcca
     2521 cccagtccaa gctgtccaaa gaccccaacg agaagcgcga tcacatggtc cttctggaat
     2581 tcgtgaccgc cgccgggatc actcacggca tggacgagct gtacaagtaa gcggccgcga
     2641 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2701 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2881 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2941 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3001 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3061 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3121 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3181 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3241 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3301 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3361 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3421 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3481 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3541 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3661 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3721 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3781 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3841 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3901 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3961 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4021 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4081 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4141 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4201 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4261 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4321 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4381 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4441 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4501 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4561 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4621 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4681 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4741 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4801 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4861 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4921 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4981 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5041 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5101 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5161 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5401 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5941 gataacc
//
