LOCUS       sequence-495312-        6583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SNAP-Rab6a (WT) in mammalian cells. Generated from Addgene
            #46781 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 15, 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
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..51)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    40..264
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      310..738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(397..416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     607..628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(747..767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        752..1081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      932..1067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     994..1013
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1148..1942
                     /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(1202..1221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1812..1831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2116..2237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2153..2172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2207..2226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2286..2302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2286..2302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2299..2321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2310..2326
                     /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(2334..2340,2341..2358,2359..2364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2379..2400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2517..2534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2688..3276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2768..2787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3447..4238,4239..4307))
                     /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     4070..4089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4308..4412)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4785..4996
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4947..4967
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4993..5017
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5157..5289
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5272..5291
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5334..5352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5364..5909
                     /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"
     promoter        complement(join(6579..6583,1..14))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(join(6580..6583,1..14))
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
       61 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
      121 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
      181 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
      241 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
      301 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
      361 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      421 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      481 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      541 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
      601 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
      661 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
      721 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
      781 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
      841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      901 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
      961 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1021 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     1081 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     1141 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     1201 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     1261 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     1321 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     1381 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     1441 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     1501 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     1561 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     1621 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     1681 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     1741 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     1801 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     1861 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     1921 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     1981 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2041 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2101 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2161 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2221 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     2281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2401 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2521 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2881 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3121 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3601 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4441 gtcgacggat cgggagatct tcaatattgg ccattagcca tattattcat tggttatata
     4501 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     4561 atttatattg gctcatgtcc aatatgaccg ccatgttggc attgattatt gactagttat
     4621 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     4681 taacttacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
     4741 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     4801 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4861 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4921 acaactgcga tcgcccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4981 ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt
     5041 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag
     5101 ctgcagtgac tctcttaagg tagccttgca gaagttggtc gtgaggcact gggcaggtaa
     5161 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga
     5221 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc
     5281 tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt acttaatacg
     5341 actcactata ggctagcctc gacatggaca aagactgcga aatgaagcgc accaccctgg
     5401 atagccctct gggcaagctg gaactgtctg ggtgcgaaca gggcctgcac cgtatcatct
     5461 tcctgggcaa aggaacatct gccgccgacg ccgtggaagt gcctgcccca gccgccgtgc
     5521 tgggcggacc agagccactg atgcaggcca ccgcctggct caacgcctac tttcaccagc
     5581 ctgaggccat cgaggagttc cctgtgccag ccctgcacca cccagtgttc cagcaggaga
     5641 gctttacccg ccaggtgctg tggaaactgc tgaaagtggt gaagttcgga gaggtcatca
     5701 gctacagcca cctggccgcc ctggccggca atcccgccgc caccgccgcc gtgaaaaccg
     5761 ccctgagcgg aaatcccgtg cccattctga tcccctgcca ccgggtggtg cagggcgacc
     5821 tggacgtggg gggctacgag ggcgggctcg ccgtgaaaga gtggctgctg gcccacgagg
     5881 gccacagact gggcaagcct gggctgggtt ctagaggatc caccatgtcc acgggcggag
     5941 acttcgggaa tccgctgagg aaattcaagc tggtgttcct gggggagcaa agcgttggaa
     6001 agacatcttt gatcaccaga ttcatgtatg acagttttga caacacctat caggcaacaa
     6061 ttggcattga ctttttatca aaaactatgt acttggagga tcgaacagta cgattgcaat
     6121 tatgggacac agcaggtcaa gagcggttca ggagcttgat tcctagctac attcgtgact
     6181 ccactgtggc agttgttgtt tatgatatca caaatgttaa ctcattccag caaactacaa
     6241 agtggattga tgatgtcaga acagaaagag gaagtgatgt tatcatcatg ctagtaggaa
     6301 ataaaacaga tcttgctgac aagaggcaag tgtcaattga ggagggagag aggaaagcca
     6361 aagagctgaa tgttatgttt attgaaacta gtgcaaaagc tggatacaat gtaaagcagc
     6421 tctttcgacg tgtagcagca gctttgccgg gaatggaaag cacacaggac agaagcagag
     6481 aagatatgat tgacataaaa ctggaaaagc ctcaggagca accagtcagt gaaggaggct
     6541 gttcctgcta actcgagcat gcatctagag ggccctattc tat
//
