LOCUS       sequence-235295-        6657 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 6657)
  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 6657)
  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: mCherry-TMCC1
FEATURES             Location/Qualifiers
     source          1..6657
                     /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"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             249..956
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(394..412)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     662..681
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3081..3202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3118..3137)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3172..3191
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3209..3664)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3346..3367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3558..3577
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3691..3795
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3797..4154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3797..3988
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3820..3840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4005..4140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4189..4983
                     /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             4189..4983
                     /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(4243..4262)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4853..4872
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5171..5190)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5215..5262
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5591..6179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6080..6099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6354..6657
                     /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 gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
      301 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
      361 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
      421 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
      481 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
      541 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
      601 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
      661 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
      721 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
      781 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
      841 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
      901 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtccg
      961 gactcagatc tcgagctatg gagccttcgg gcagtgaaca gttatttgag gaccctgatc
     1021 ctggaggcaa atcccaagat gcagaggcca gaaagcagac agaatcagaa caaaaattgt
     1081 ctaaaatgac ccacaatgct ttggagaaca ttaacgtgat tggccaaggc ttgaagcatc
     1141 tcttccagca ccagcgcagg aggtcatcag tgtctccaca tgatgtgcag caaattcagg
     1201 cagatccaga acctgaaatg gatctggaaa gccagaacgc atgtgctgag attgatggtg
     1261 tccccaccca ccccacagct ctgaatcgtg tcctgcagca gattcgagtg ccacccaaga
     1321 tgaagagagg gacaagcttg catagtaggc ggggcaagcc agaggcccca aagggaagtc
     1381 cccaaatcaa caggaagtct ggtcaggaga tgacagctgt tatgcagtca ggccgaccca
     1441 ggtcttcatc cacaactgat gcacctacca gctctgctat gatggaaata gcttgtgctg
     1501 ctgctgctgc tgctgctgca tgtctaccag gagaggaggg aactgcggag cggatcgaac
     1561 ggttggaagt aagcagcctt gcccaaacat ccagtgcagt ggcctccagt accgatggca
     1621 gcatccacac agactctgtg gatggaacac cagaccctca gcgcacaaag gctgccattg
     1681 ctcacctgca gcagaagatc ctgaagctca cagaacaaat caagattgca caaacagccc
     1741 gggacgacaa cgttgctgaa tacttgaagc ttgccaacag tgcagacaaa cagcaggctg
     1801 cccgcatcaa gcaagtcttt gagaagaaga accagaaatc tgcccaaact atcctccagc
     1861 tgcaaaagaa acttgagcac taccacagga agctcagaga ggtagagcag aatgggatcc
     1921 cccggcagcc aaaggatgtc ttcagggaca tgcaccaggg tctgaaggat gtaggagcaa
     1981 aggtgactgg cttcagtgaa ggtgtggtgg atagtgtcaa aggtgggttt tccagcttct
     2041 cccaggccac ccattcagca gcaggcgctg tagtctcaaa gcccagagag attgcctcac
     2101 tcattcggaa caaatttggc agtgcagaca acatccccaa cctgaaggac tctttagagg
     2161 aagggcaagt ggatgatgcg gggaaggctt tgggagtgat ttcaaacttt cagtctagcc
     2221 caaaatatgg tagtgaagaa gattgttcta gtgccacttc aggctcagtg ggagccaaca
     2281 gcaccacagg gggcatcgct gtaggagcat ccagctccaa aacaaacacc ctggacatgc
     2341 agagctcagg atttgatgca ctactacatg agatccagga gatccgggaa acccaggcca
     2401 gactagagga atcctttgag actctcaagg aacattatca gagggactat tccttaataa
     2461 tgcagacctt acaggaggag cgatatagat gtgaacgatt ggaagaacag ctaaatgacc
     2521 taacagagct ccaccagaat gaaatcttga acttgaagca ggaactggca agcatggaag
     2581 aaaaaatcgc gtatcagtcc tatgaacggg cccgggacat ccaggaggcc ctggaggcat
     2641 gccagacgcg catctccaag atggagctgc agcagcagca gcagcaggtg gtgcagctag
     2701 aagggctgga gaatgccact gcccggaacc ttctgggcaa actcatcaac atcctcctgg
     2761 ctgtcatggc agtccttttg gtctttgtct ccactgtagc caactgtgtg gtccccctca
     2821 tgaagactcg caacaggacg ttcagcactt tattccttgt ggtttttatt gcctttctct
     2881 ggaagcactg ggacgccctc ttcagctatg tggaacggtt cttttcatcc cctagaggta
     2941 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     3001 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3061 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3121 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3181 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     3241 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3301 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3361 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3421 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3481 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3541 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3601 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3661 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3721 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3781 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3901 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3961 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4021 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4081 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4141 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4201 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4261 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4321 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4381 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4441 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4501 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4561 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4621 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4681 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4741 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4801 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4861 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4921 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4981 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5041 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5101 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5161 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5221 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5281 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5341 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5401 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5461 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5521 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5581 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5641 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5701 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5761 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5821 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5881 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5941 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6001 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6061 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6121 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6181 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6241 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     6301 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6361 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6421 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6481 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6541 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6601 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatg
//
