LOCUS       sequence-235373-        6849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human TMCC2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6849)
  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 6849)
  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: GFP-TMCC2
FEATURES             Location/Qualifiers
     source          1..6849
                     /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..965
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    3273..3394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3310..3329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3364..3383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3401..3856)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3538..3559)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3750..3769
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3883..3987
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3989..4346
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3989..4180
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4012..4032)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4197..4332
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4381..5175
                     /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             4381..5175
                     /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(4435..4454)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5045..5064
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5363..5382)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5407..5454
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5783..6371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6272..6291
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6546..6849
                     /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 ggcgccgagc tgttcaccgg catcgtgccc atcctgatcg
      301 agctgaatgg cgatgtgaat ggccacaagt tcagcgtgag cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cctgtgccct
      421 ggcccaccct ggtgaccacc ctgagctacg gcgtgcagtg cttctcacgc taccccgatc
      481 acatgaagca gcacgacttc ttcaagagcg ccatgcctga gggctacatc caggagcgca
      541 ccatcttctt cgaggatgac ggcaactaca agtcgcgcgc cgaggtgaag ttcgagggcg
      601 ataccctggt gaatcgcatc gagctgaccg gcaccgattt caaggaggat ggcaacatcc
      661 tgggcaataa gatggagtac aactacaacg cccacaatgt gtacatcatg accgacaagg
      721 ccaagaatgg catcaaggtg aacttcaaga tccgccacaa catcgaggat ggcagcgtgc
      781 agctggccga ccactaccag cagaataccc ccatcggcga tggccctgtg ctgctgcccg
      841 ataaccacta cctgtccacc cagagcgccc tgtccaagga ccccaacgag aagcgcgatc
      901 acatgatcta cttcggcttc gtgaccgccg ccgccatcac ccacggcatg gatgagctgt
      961 acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccatga
     1021 agaggtgcag atcggacgag ctgcagcaac aacagggcga ggaggatgga gctgggctgg
     1081 aagatgccgc ttcccacctg ccgggcgcgg acctccggcc tggggagacc acgggtgcta
     1141 actctgctgg cgggccaact tcagacgccg gcgctgccgc ggcgcccaac ccaggtcccc
     1201 gaagcaagcc tcctgattta aagaaaatcc agcagctgtc agagggctcc atgtttggcc
     1261 acggtctgaa gcacctgttc cacagccgcc gtcggtctcg ggaaagggag caccagacgt
     1321 ctcaggattc ccagcagcat cagcagcagc agggtatgtc cgaccatgac tccccagatg
     1381 agaaggagcg ctctccggag atgcatcgcg tctcctacgc catgtccctg cacgacctgc
     1441 ccgcccggcc caccgccttc aaccgcgtgc tgcagcagat ccgctcccgg ccctccatca
     1501 agcggggcgc cagcctgcac agcagcagtg ggggcggcag cagcgggagc agcagccggc
     1561 gcaccaagag tagctccctg gagccccagc gtggcagccc tcacctgctg cgcaaggccc
     1621 cccaggacag cagcctggcc gccatcctgc accagcacca gtgccgtccc cgctcttcct
     1681 ccaccaccga cactgctctg ctgctggccg acggcagcaa cgtgtacctc ctggctgagg
     1741 aggccgaagg catcggggac aaggtcgata agggagacct ggtggccctg agcctccccg
     1801 ccggccatgg tgacaccgac ggccccatca gcctggacgt gcccgatggg gcaccggacc
     1861 cccagcggac caaggccgcc attgaccacc tgcaccagaa gatcctgaag atcaccgagc
     1921 agatcaagat tgagcaggag gctcgcgacg acaatgtggc agagtatctg aaactggcca
     1981 acaacgcgga caagcagcag gtgtcacgca tcaagcaagt gttcgagaag aagaaccaga
     2041 agtcagccca gaccatcgcc cagctgcaca agaagctgga gcactaccgc cggcgcctga
     2101 aggagattga gcagaacggg ccctcgcggc agcccaagga cgtgctgcgg gacatgcagc
     2161 aggggctgaa ggacgtgggc gccaacgtgc gcgcaggcat cagcggcttt gggggcggcg
     2221 tggtggaggg cgtcaagggc agcctctctg gcctctcaca ggccacccac accgccgtgg
     2281 tgtccaagcc ccgggagttt gccagcctca tccgcaacaa gtttggcagt gctgacaaca
     2341 tcgcccacct gaaggacccc ctggaagatg ggccccctga ggaggcagcc cgggcactga
     2401 gcggcagtgc cacactcgtc tccagcccca agtatggcag cgatgatgag tgctccagcg
     2461 ccagcgccag ctcagccggg gcaggcagca actctggggc tgggcctggt ggggcgctgg
     2521 ggagccctaa gtccaatgca ctgtatggtg ctcctggaaa cctggatgct ctgctggaag
     2581 agctacggga gatcaaggag ggacagtctc acctggagga ctccatggaa gacctgaaga
     2641 ctcagctgca gagggactac acctacatga cccagtgcct gcaggaggag cgctacaggt
     2701 acgagcggct ggaggagcag ctcaacgacc tgactgagct tcatcagaac gagatgacga
     2761 acctgaagca ggagctggcc agcatggagg agaaggtggc ctaccagtcc tatgagaggg
     2821 cacgggacat ccaggaggcc gtggagtcct gcctgacccg ggtcaccaag ctggagctgc
     2881 agcagcaaca gcagcaggtg gtacagctgg agggcgtgga gaatgccaac gcgcgggcgc
     2941 tgctgggcaa gttcatcaac gtgatcctgg cgctcatggc cgtgctgctg gtgttcgtgt
     3001 ccaccatcgc caacttcatc acgcccctca tgaagacacg cctgcgcatc accagcacca
     3061 ccctcctggt cctcgtcctg ttcctcctct ggaagcactg ggactccctc acctacctcc
     3121 tggagcacgt gttgctgccc agcggatcca ccggatctag ataactgatc ataatcagcc
     3181 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3241 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3301 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3361 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     3421 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3481 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3541 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3601 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3661 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3721 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3781 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3841 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3901 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3961 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4021 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4141 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4201 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4261 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4321 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4381 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4441 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4501 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4561 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4621 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4681 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4741 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4801 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4861 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4921 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4981 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5041 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5101 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5161 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5221 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5281 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5341 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5401 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5461 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5521 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5581 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6241 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6301 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6361 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6421 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6481 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     6541 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     6601 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     6661 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     6721 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     6781 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6841 attaccatg
//
