LOCUS       sequence-235377-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human TMCC1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Hoyer MJ, Chitwood PJ, Ebmeier CC, Striepen JF, Qi RZ, Old WM, 
            Voeltz GK
  TITLE     A Novel Class of ER Membrane Proteins Regulates ER-Associated 
            Endosome Fission.
  JOURNAL   Cell. 2018 Sep 20;175(1):254-265.e14. doi: 
            10.1016/j.cell.2018.08.030. Epub 2018 Sep 13.
  PUBMED    30220460
REFERENCE   2  (bases 1 to 5993)
  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"; doi: 
            "10.1016/j.cell.2018.08.030"; journalName: "Cell"; date: 
            "2018-09-20- 20"; volume: "175"; issue: "1"; pages: "254-265.e14"
COMMENT     Sequence Label: 3XFlag-TMCC1
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             236..301
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2417..2538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2454..2473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2508..2527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2545..3000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2682..2703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2894..2913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3027..3131
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3133..3490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3133..3324
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3156..3176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3341..3476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3525..4319
                     /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             3525..4319
                     /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(3579..3598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4189..4208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4507..4526)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4551..4598
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4927..5515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5416..5435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5690..5993
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatggacta
      241 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
      301 gaaagcttct cgagctgagc cttcgggcag tgaacagtta tttgaggacc ctgatcctgg
      361 aggcaaatcc caagatgcag aggccagaaa gcagacagaa tcagaacaaa aattgtctaa
      421 aatgacccac aatgctttgg agaacattaa cgtgattggc caaggcttga agcatctctt
      481 ccagcaccag cgcaggaggt catcagtgtc tccacatgat gtgcagcaaa ttcaggcaga
      541 tccagaacct gaaatggatc tggaaagcca gaacgcatgt gctgagattg atggtgtccc
      601 cacccacccc acagctctga atcgtgtcct gcagcagatt cgagtgccac ccaagatgaa
      661 gagagggaca agcttgcata gtaggcgggg caagccagag gccccaaagg gaagtcccca
      721 aatcaacagg aagtctggtc aggagatgac agctgttatg cagtcaggcc gacccaggtc
      781 ttcatccaca actgatgcac ctaccagctc tgctatgatg gaaatagctt gtgctgctgc
      841 tgctgctgct gctgcatgtc taccaggaga ggagggaact gcggagcgga tcgaacggtt
      901 ggaagtaagc agccttgccc aaacatccag tgcagtggcc tccagtaccg atggcagcat
      961 ccacacagac tctgtggatg gaacaccaga ccctcagcgc acaaaggctg ccattgctca
     1021 cctgcagcag aagatcctga agctcacaga acaaatcaag attgcacaaa cagcccggga
     1081 cgacaacgtt gctgaatact tgaagcttgc caacagtgca gacaaacagc aggctgcccg
     1141 catcaagcaa gtctttgaga agaagaacca gaaatctgcc caaactatcc tccagctgca
     1201 aaagaaactt gagcactacc acaggaagct cagagaggta gagcagaatg ggatcccccg
     1261 gcagccaaag gatgtcttca gggacatgca ccagggtctg aaggatgtag gagcaaaggt
     1321 gactggcttc agtgaaggtg tggtggatag tgtcaaaggt gggttttcca gcttctccca
     1381 ggccacccat tcagcagcag gcgctgtagt ctcaaagccc agagagattg cctcactcat
     1441 tcggaacaaa tttggcagtg cagacaacat ccccaacctg aaggactctt tagaggaagg
     1501 gcaagtggat gatgcgggga aggctttggg agtgatttca aactttcagt ctagcccaaa
     1561 atatggtagt gaagaagatt gttctagtgc cacttcaggc tcagtgggag ccaacagcac
     1621 cacagggggc atcgctgtag gagcatccag ctccaaaaca aacaccctgg acatgcagag
     1681 ctcaggattt gatgcactac tacatgagat ccaggagatc cgggaaaccc aggccagact
     1741 agaggaatcc tttgagactc tcaaggaaca ttatcagagg gactattcct taataatgca
     1801 gaccttacag gaggagcgat atagatgtga acgattggaa gaacagctaa atgacctaac
     1861 agagctccac cagaatgaaa tcttgaactt gaagcaggaa ctggcaagca tggaagaaaa
     1921 aatcgcgtat cagtcctatg aacgggcccg ggacatccag gaggccctgg aggcatgcca
     1981 gacgcgcatc tccaagatgg agctgcagca gcagcagcag caggtggtgc agctagaagg
     2041 gctggagaat gccactgccc ggaaccttct gggcaaactc atcaacatcc tcctggctgt
     2101 catggcagtc cttttggtct ttgtctccac tgtagccaac tgtgtggtcc ccctcatgaa
     2161 gactcgcaac aggacgttca gcactttatt ccttgtggtt tttattgcct ttctctggaa
     2221 gcactgggac gccctcttca gctatgtgga acggttcttt tcatccccta gaggtaccgc
     2281 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     2341 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2401 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2521 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2581 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2641 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2701 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2761 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2821 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2881 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2941 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3001 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3061 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3121 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3421 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3481 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3541 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3601 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3661 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3721 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3781 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3841 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3901 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3961 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4021 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4081 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4141 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4201 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4261 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4321 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4381 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4441 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4501 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4561 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4621 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4681 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4741 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4801 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4861 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4921 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4981 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5041 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5101 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5161 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5221 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5281 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5341 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5401 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5461 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5521 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5581 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     5641 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     5701 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     5761 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     5821 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     5881 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     5941 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atg
//
