LOCUS       sequence-487506-        5274 bp    DNA     circular SYN 30-SEP-2025
DEFINITION  Split Apex4 works with Nterm Apex human wild-type Rab27a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5274)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 5274)
  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: Nterm Apex4 3xFlag Rab27a
FEATURES             Location/Qualifiers
     source          1..5274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1204..1269
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2074..2195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2111..2130)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2165..2184
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2202..2657)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2339..2360)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2551..2570
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2684..2788
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2813..2833)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2857..3067
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2879..3075
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2926..3061
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3110..3904
                     /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             3110..3904
                     /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(3164..3183)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3774..3793
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4092..4111)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4136..4183
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4512..5100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5001..5020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggggaa aagttatccc actgtatcag cggactacca ggacgctgtg
      601 gagaaggcta agaagagact cggaggattt atcgctgaaa aacgatgcgc tccgctcatg
      661 ttgcgacttg cattccactc cgctggtaca ttcgataagc gaactaagac tggcggaccc
      721 tttgggacaa ttagataccc cgccgagctt gcgcacagtg ccaattctgg cttggatata
      781 gcggtacggc tccttgagcc ccttaaagca gagttcccta tccttagtta cgctgacttc
      841 taccagctcg cgggcgtagt agctgtcgag gtgacgggcg ggcctaaagt gccctttcat
      901 ccgggaagag aggacaagcc ggaacttcca cccgaaggac gacttccaga cccaaccaaa
      961 gggagtgatc atctccggga cgtatttggc aaagctatgg ggctgactga tcaggacata
     1021 gtagccctct caggtggcca taccttgggt gctgcacata aagagagaag tggatttgag
     1081 ggtccctgga cgagtaatcc attggtattc gataactcct actttactga acttctttcc
     1141 ggtgagaagg aaggctcggg cagcacctcg ggcagcggct cgacctcggg ctcgggctcc
     1201 ggagactaca aggaccatga cggtgactat aaggaccacg acatcgacta caaagatgat
     1261 gatgataagg gctcgagtat gtctgatgga gattatgatt acctcatcaa gtttttagct
     1321 ttgggagact ctggtgtagg gaagaccagt gtactttacc aatatacaga tggtaaattt
     1381 aactccaaat ttatcacaac agtgggcatt gatttcaggg aaaaaagagt ggtgtacaga
     1441 gccagtgggc cggatggagc cactggcaga ggccagagaa tccacctgca gttatgggac
     1501 acagcagggc aggagaggtt tcgtagctta acgacagcgt tcttcagaga tgctatgggt
     1561 tttcttctac tttttgatct gacaaatgag caaagtttcc tcaatgtcag aaactggata
     1621 agccagctac agatgcatgc atattgtgaa aacccagata tagtgctgtg tggaaacaag
     1681 agtgatctgg aggaccagag agtagtgaaa gaggaggaag ccatagcact cgcagagaaa
     1741 tatggaatcc cctactttga aactagtgct gccaatggga caaacataag ccaagcaatt
     1801 gagatgcttc tggacctgat aatgaagcga atggaacggt gtgtggacaa gtcctggatt
     1861 cctgaaggag tggtgcgatc aaatggtcat gcctctacgg atcagttaag tgaagaaaag
     1921 gagaaagggg catgtggctg ttaaggatcc accggatcta gataactgat cataatcagc
     1981 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2041 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2101 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2161 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     2221 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2281 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2341 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2401 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2461 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2521 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2581 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2641 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2701 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2761 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     2821 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2881 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2941 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3001 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3061 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3121 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3181 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3241 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3301 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3361 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3421 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3481 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3541 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3601 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     3661 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3721 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3781 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3841 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3901 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3961 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4021 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4081 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4141 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4201 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4261 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4321 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4501 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4561 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4621 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4681 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4741 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4801 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4861 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4921 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4981 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5041 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5101 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5161 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     5221 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
