LOCUS       Exported                6153 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses human TMCC3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    GFP-TMCC3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6153)
  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 6153)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6153
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             249..965
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2577..2698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2614..2633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2668..2687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2705..3160)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2842..2863)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3054..3073
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3187..3291
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3293..3650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3316..3336)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3501..3636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3563..3582
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3685..4479
                     /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(3739..3758)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4349..4368
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4667..4686)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4711..4758
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5087..5675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5576..5595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5850..6153
                     /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 ggtaccatgc
     1021 cgggcagcga cacggcgctc accgtggacc ggacctactc gtaccccggc cggcaccacc
     1081 gctgcaagag ccgggtagaa cgtcatgaca tgaatacctt aagcctgccc ctgaacatac
     1141 gccgaggggg gtcagacacc aacctcaact ttgatgtccc ggatggcatc ctggacttcc
     1201 acaaggtcaa actcactgca gacagcctga agcaaaaaat tctaaaggta acagagcaga
     1261 taaaaattga gcaaacatcg cgcgatggga atgttgcgga gtatctgaaa ctagtgaaca
     1321 acgcggacaa gcagcaggcg ggacgtatca agcaagtctt tgagaagaag aatcagaaat
     1381 cagctcactc catcgcccag ctgcagaaga agttagagca gtatcatcga aagctcagag
     1441 agatcgagca gaatggagcc tctaggagct caaaggacat ttccaaagac cacctgaagg
     1501 atatacatcg ctctttgaaa gatgcccacg tgaaatctcg aactgccccc cattgcatgg
     1561 agagcagcaa atcgggcatg ccaggggtct cacttactcc acctgtgttt gttttcaata
     1621 agtccagaga gtttgccaac ctgatccgga ataagtttgg cagcgccgac aacattgctc
     1681 acttgaaaaa ttccttagag gagtttaggc cagaggcgag tgccagggcc tacgggggca
     1741 gcgctaccat cgtgaacaaa cccaagtatg gcagtgatga tgaatgttcg agtggcacgt
     1801 caggctcggc cgacagtaac ggaaaccagt cgtttggggc tggtggagcc agcacactgg
     1861 acagccaggg caagctcgcc gtgatcctgg aggaactgag ggagatcaag gatacccaag
     1921 ctcagctggc tgaggacatc gaggcactga aggtgcagtt taagagagaa tatggtttta
     1981 tttctcagac cctgcaagag gaaagataca ggtatgagcg actggaggac cagctgcatg
     2041 acctgacgga cctgcatcag catgagacag ccaacctgaa gcaggagctg gccagcattg
     2101 aggagaaggt ggcctaccag gcctacgagc gctcgcggga catccaggaa gccttggaat
     2161 cctgccagac tcgcatttct aagctggagc tccaccagca agagcagcaa gctctgcaga
     2221 cagacaccgt gaatgctaaa gttctcctgg ggaggtgcat caacgtgatc ctggccttca
     2281 tgactgtcat cttagtgtgt gtgtccacca tcgcgaagtt cgtctcaccc atgatgaaga
     2341 gtcgctgcca cattcttggc accttctttg ccgtgactct tcttgctata ttttgtaaaa
     2401 actgggacca tatcctgtgt gccatagaaa ggatgataat accaagagga tccaccggat
     2461 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2521 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2581 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2641 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     2701 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2761 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2821 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2881 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2941 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3001 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3061 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3121 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3181 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3241 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3301 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3481 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3541 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3601 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3661 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3721 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3781 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3841 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3901 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3961 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4021 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4081 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4141 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4201 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4261 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4321 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4381 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4441 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4501 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4561 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4621 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4681 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4741 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4801 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4861 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4921 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4981 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5221 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5281 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5341 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5401 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5461 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5521 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5581 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5641 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5701 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5761 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     5821 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     5881 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5941 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     6001 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     6061 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6121 tacatctacg tattagtcat cgctattacc atg
//
