LOCUS       sequence-487483-        5801 bp    DNA     circular SYN 30-SEP-2025
DEFINITION  Distribution and localization of human wild-type Rab27a in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5801)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 5801)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mCherry Rab27a
FEATURES             Location/Qualifiers
     source          1..5801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    32..52
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        113..416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        417..620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    641..665
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      666..675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             672..1379
                     /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(817..835)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1085..1104
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2046..2070)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2099..2220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2110..2129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2164..2183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     enhancer        2318..2697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2698..2901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2934..3533
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2934..2953)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3430..3450
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3571..3588)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3577..3801
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3826..3843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3997..4585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4077..4096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4756..5547,5548..5616))
                     /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     5379..5398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5617..5721)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5789..5801,1..6)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgaggccctt tcgtcggcgc gccgcggccg ccaactttgt atagaaaagt tgtagttatt
       61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      601 aggtctatat aagcagagct ggtttagtga accgtcagat ccaagtttgt acaaaaaagc
      661 aggctgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
      721 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      781 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      841 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
      901 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      961 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     1021 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1081 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1141 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1201 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1261 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1321 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga
     1381 tgtctgatgg agattatgat tacctcatca agtttttagc tttgggagac tctggtgtag
     1441 ggaagaccag tgtactttac caatatacag atggtaaatt taactccaaa tttatcacaa
     1501 cagtgggcat tgatttcagg gaaaaaagag tggtgtacag agccagtggg ccggatggag
     1561 ccactggcag aggccagaga atccacctgc agttatggga cacagcaggg caggagaggt
     1621 ttcgtagctt aacgacagcg ttcttcagag atgctatggg ttttcttcta ctttttgatc
     1681 tgacaaatga gcaaagtttc ctcaatgtca gaaactggat aagccagcta cagatgcatg
     1741 catattgtga aaacccagat atagtgctgt gtggaaacaa gagtgatctg gaggaccaga
     1801 gagtagtgaa agaggaggaa gccatagcac tcgcagagaa atatggaatc ccctactttg
     1861 aaactagtgc tgccaatggg acaaacataa gccaagcaat tgagatgctt ctggacctga
     1921 taatgaagcg aatggaacgg tgtgtggaca agtcctggat tcctgaagga gtggtgcgat
     1981 caaatggtca tgcctctacg gatcagttaa gtgaagaaaa ggagaaaggg gcatgtggct
     2041 gttgaaccca gctttcttgt acaaagtggt gatggccggc cgcttcgagc agacatgata
     2101 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2161 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2221 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2281 taaagcaagt aaaacctcta caaatgtggt acgcgttgac attgattatt gactagttat
     2341 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2401 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2461 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2521 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2581 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2641 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2701 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2761 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2821 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2881 gaggtctata taagcagagc tctctggcta actagagaac ccactgcgcc accatgaccg
     2941 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     3001 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     3061 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     3121 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     3181 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     3241 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     3301 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     3361 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     3421 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     3481 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgactcgagt
     3541 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3601 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3661 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3721 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3781 gggatgcggt gggctctatg ggcggccgcg gcgctcttcc gcttcctcgc tcactgactc
     3841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4141 taccggatac ctgtccgcct ttctctcttc gggaagcgtg gcgctttctc atagctcacg
     4201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5761 cattattatc atgacattaa cctataaaaa taggcgtatc a
//
