LOCUS       sequence-495397-        6583 bp    DNA     circular SYN 16-DEC-2025
DEFINITION  Expresses SNAP-Rab6a-T27N (dominant negative) in mammalian cells. 
            Generated from Addgene #46782 plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  AUTHORS   Hekmatara M, Thompson SL, Haggerty KN, Hagen S, Brothers BA, Daniels
            B, Luxardi G, Moshiri A, Deng WT, Robichaux MA
  TITLE     Super-resolution microscopy reveals a Rab6a-dependent trafficking 
            hub for rhodopsin at the mammalian rod photoreceptor Golgi.
  JOURNAL   bioRxiv [Preprint]. 2025 Sep 25:2025.09.24.678292. doi: 
            10.1101/2025.09.24.678292.
  PUBMED    41040232
REFERENCE   2  (bases 1 to 6583)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Sep 25:2025.09.24.678292. doi: 
            10.1101/2025.09.24.678292."
COMMENT     Sequence Label: pCMV-SNAP-Rab6a_T27N
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(892..910)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(893..910)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(930..947)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    936..1160
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1206..1634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1293..1312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1503..1524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1643..1663)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1648..1977
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1828..1963
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1890..1909
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2044..2838
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2098..2117)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2708..2727
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3012..3133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3049..3068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3103..3122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3182..3198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3182..3198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3195..3217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3206..3222
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3230..3236,3237..3254,3255..3260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3275..3296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3413..3430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3584..4172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3664..3683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4343..5134,5135..5203))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4966..4985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5204..5308)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5681..5892
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5843..5863
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5889..5913
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6053..6185
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6168..6187
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        6230..6248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(6260..6583,1..222)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
ORIGIN
        1 ggagaggtca tcagctacag ccacctggcc gccctggccg gcaatcccgc cgccaccgcc
       61 gccgtgaaaa ccgccctgag cggaaatccc gtgcccattc tgatcccctg ccaccgggtg
      121 gtgcagggcg acctggacgt ggggggctac gagggcgggc tcgccgtgaa agagtggctg
      181 ctggcccacg agggccacag actgggcaag cctgggctgg gttctagagg atccaccatg
      241 tccacgggcg gagacttcgg gaatccgctg aggaaattca agctggtgtt cctgggggag
      301 caaagcgttg gaaagaattc tttgatcacc agattcatgt atgacagttt tgacaacacc
      361 tatcaggcaa caattggcat tgacttttta tcaaaaacta tgtacttgga ggatcgaaca
      421 gtacgattgc aattatggga cacagcaggt caagagcggt tcaggagctt gattcctagc
      481 tacattcgtg actccactgt ggcagttgtt gtttatgata tcacaaatgt taactcattc
      541 cagcaaacta caaagtggat tgatgatgtc agaacagaaa gaggaagtga tgttatcatc
      601 atgctagtag gaaataaaac agatcttgct gacaagaggc aagtgtcaat tgaggaggga
      661 gagaggaaag ccaaagagct gaatgttatg tttattgaaa ctagtgcaaa agctggatac
      721 aatgtaaagc agctctttcg acgtgtagca gcagctttgc cgggaatgga aagcacacag
      781 gacagaagca gagaagatat gattgacata aaactggaaa agcctcagga gcaaccagtc
      841 agtgaaggag gctgttcctg ctaactcgag catgcatcta gagggcccta ttctatagtg
      901 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
      961 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1021 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1081 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1141 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     1201 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1261 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1321 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1381 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1441 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1501 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1561 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1621 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     1681 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     1741 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1801 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     1861 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     1921 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     1981 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     2041 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2101 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2161 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2221 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2281 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2341 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2401 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2461 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2521 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     2581 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2641 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     2701 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     2761 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     2821 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     2881 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     2941 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3001 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3061 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3121 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3181 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3241 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3301 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3361 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4021 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4261 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4321 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4381 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4441 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4501 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4561 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4621 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4681 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4741 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4801 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4861 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4921 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4981 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5041 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5101 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5161 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5221 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5281 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     5341 acggatcggg agatcttcaa tattggccat tagccatatt attcattggt tatatagcat
     5401 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     5461 atattggctc atgtccaata tgaccgccat gttggcattg attattgact agttattaat
     5521 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     5581 ttacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     5641 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     5701 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     5761 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     5821 ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     5881 ataagcagag ctcgtttagt gaaccgtcag atcactagaa gctttattgc ggtagtttat
     5941 cacagttaaa ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa cttaagctgc
     6001 agtgactctc ttaaggtagc cttgcagaag ttggtcgtga ggcactgggc aggtaagtat
     6061 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     6121 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     6181 cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc
     6241 actataggct agcctcgaca tggacaaaga ctgcgaaatg aagcgcacca ccctggatag
     6301 ccctctgggc aagctggaac tgtctgggtg cgaacagggc ctgcaccgta tcatcttcct
     6361 gggcaaagga acatctgccg ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg
     6421 cggaccagag ccactgatgc aggccaccgc ctggctcaac gcctactttc accagcctga
     6481 ggccatcgag gagttccctg tgccagccct gcaccaccca gtgttccagc aggagagctt
     6541 tacccgccag gtgctgtgga aactgctgaa agtggtgaag ttc
//
