LOCUS       sequence-322819-        6793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR mutant with truncated cytosolic tail and TMD in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  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 6793)
  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: F9F17F-GFP
FEATURES             Location/Qualifiers
     source          1..6793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      630..639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1389..1400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2745..2754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2751..2753,2754..2756,2757..3470)
                     /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(2796..2817)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3592..3713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3629..3648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3683..3702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3720..4175)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3857..3878)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4069..4088
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4202..4306
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4308..4665
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4308..4499
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4331..4351)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4516..4651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4700..5494
                     /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             4700..5494
                     /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(4754..4773)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5364..5383
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5682..5701)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5726..5773
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6102..6690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6591..6610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggactca gatctcgagg ccaccatggc caatgtcaca aaaccaaaaa
      661 gggctgtgat cgtctttttc ttgattggat ttatgattgg ctacttgggc tattgtaaag
      721 gggtagaacc aaaaactgag tgtgagagac tggcaggaac cgagtctcca gtgagggagg
      781 agccaggaga ggacttccct gcagcacgtc gcttatattg ggatgacctg aagagaaagt
      841 tgtcggagaa actggacagc acagacttca ccagcaccat caagctgctg aatgaaaatt
      901 catatgtccc tcgtgaggct ggatctcaaa aagatgaaaa tcttgcgttg tatgttgaaa
      961 atcaatttcg tgaatttaaa ctcagcaaag tctggcgtga tcaacatttt gttaagattc
     1021 aggtcaaaga cagcgctcaa aactcggtga tcatagttga taagaacggt agacttgttt
     1081 acctggtgga gaatcctggg ggttatgtgg cgtatagtaa ggctgcaaca gttactggta
     1141 aactggtcca tgctaatttt ggtactaaaa aagattttga ggatttatac actcctgtga
     1201 atggatctat agtgattgtc agagcaggga aaatcacctt tgcagaaaag gttgcaaatg
     1261 ctgaaagctt aaatgcaatt ggtgtgttga tatacatgga ccagactaaa tttcccattg
     1321 ttaacgcaga actttcattc tttggacatg ctcatctggg gacaggtgac ccttacacac
     1381 ctggattccc ttccttcaat cacactcagt ttccaccatc tcggtcatca ggattgccta
     1441 atatacctgt ccagacaatc tccagagctg ctgcagaaaa gctgtttggg aatatggaag
     1501 gagactgtcc ctctgactgg aaaacagact ctacatgtag gatggtaacc tcagaaagca
     1561 agaatgtgaa gctcactgtg agcaatgtgc tgaaagagat aaaaattctt aacatctttg
     1621 gagttattaa aggctttgta gaaccagatc actatgttgt agttggggcc cagagagatg
     1681 catggggccc tggagctgca aaatccggtg taggcacagc tctcctattg aaacttgccc
     1741 agatgttctc agatatggtc ttaaaagatg ggtttcagcc cagcagaagc attatctttg
     1801 ccagttggag tgctggagac tttggatcgg ttggtgccac tgaatggcta gagggatacc
     1861 tttcgtccct gcatttaaag gctttcactt atattaatct ggataaagcg gttcttggta
     1921 ccagcaactt caaggtttct gccagcccac tgttgtatac gcttattgag aaaacaatgc
     1981 aaaatgtgaa gcatccggtt actgggcaat ttctatatca ggacagcaac tgggccagca
     2041 aagttgagaa actcacttta gacaatgctg ctttcccttt ccttgcatat tctggaatcc
     2101 cagcagtttc tttctgtttt tgcgaggaca cagattatcc ttatttgggt accaccatgg
     2161 acacctataa ggaactgatt gagaggattc ctgagttgaa caaagtggca cgagcagctg
     2221 cagaggtcgc tggtcagttc gtgattaaac taacccatga tgttgaattg aacctggact
     2281 atgagaggta caacagccaa ctgctttcat ttgtgaggga tctgaaccaa tacagagcag
     2341 acataaagga aatgggcctg agtttacagt ggctgtattc tgctcgtgga gacttcttcc
     2401 gtgctacttc cagactaaca acagatttcg ggaatgctga gaaaacagac agatttgtca
     2461 tgaagaaact caatgatcgt gtcatgagag tggagtatca cttcctctct ccctacgtat
     2521 ctccaaaaga gtctcctttc cgacatgtct tctggggctc cggctctcac acgctgccag
     2581 ctttactgga gaacttgaaa ctgcgtaaac aaaataacgg tgcttttaat gaaacgctgt
     2641 tcagaaacca gttggctcta gctacttgga ctattcaggg agctgcaaat gccctctctg
     2701 gtgacgtttg ggacattgac aatgagtttc gggatccacc ggtcgccacc atggtgtcca
     2761 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2821 acggccacaa gttctccgtg cggggcgagg gcgagggcga tgccaccaac ggcaagctga
     2881 ccctgaagtt catcagcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2941 ccctgaccta cggcgtgcag agcttctccc gctaccccga ccacatgaag cgccacgact
     3001 tcttcaagag cgccatgccc gaaggctacg tccaggagcg caccatctcc ttcaaggacg
     3061 acggcaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3121 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3181 acaacttcaa ctcccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     3241 ccaacttcaa gatccgccac aacgtggagg acggctccgt gcagctcgcc gaccactacc
     3301 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgtcca
     3361 cccagtccaa gctgtccaaa gaccccaacg agaagcgcga tcacatggtc cttctggaat
     3421 tcgtgaccgc cgccgggatc actcacggca tggacgagct gtacaagtaa gcggccgcga
     3481 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3541 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3601 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3661 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3721 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3781 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3841 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3901 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3961 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4021 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4081 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4141 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4201 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4261 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     4321 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4381 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4501 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4561 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4621 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4681 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4741 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4801 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4861 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4921 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4981 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5041 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5101 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5161 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5221 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5281 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5341 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5401 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5461 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5521 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5581 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5641 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5701 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5761 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5821 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5881 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5941 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6001 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6061 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6121 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6181 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6241 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6301 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6361 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6421 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6481 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6541 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6601 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6661 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6721 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6781 gataaccgta tta
//
