LOCUS       sequence-489499-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For exogenous expression of Rab45 (RASEF) as an N-terminal fusion to
            RFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  AUTHORS   Hanna MG, Rodriguez Cruz HO, Fujise K, Wu Y, Xu CS, Pang S, Li Z, 
            Monetti M, De Camilli P
  TITLE     BLTP3A is associated with membranes of the late endocytic pathway 
            and is an effector of CASM.
  JOURNAL   EMBO J (2025) 44: 6168 - 6195
  PUBMED    39386594
REFERENCE   2  (bases 1 to 6936)
  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: "EMBO J 
            (2025) 44: 6168 - 6195"
COMMENT     Sequence Label: pmCh-C1 Rab45
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             612..1319
                     /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(757..775)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1025..1044
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3724..3845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3761..3780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3815..3834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3852..4307)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3989..4010)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4201..4220
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4334..4438
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4440..4797
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4440..4631
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4463..4483)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4648..4783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4832..5626
                     /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             4832..5626
                     /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(4886..4905)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5496..5515
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5814..5833)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5858..5905
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6234..6822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6723..6742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
      661 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      721 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      781 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
      841 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      901 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      961 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1021 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1081 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1141 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1201 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1261 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg
     1321 gtaccggtgg atctggaggt atggaggctg acggcgacgg cgaagagctg gccagactga
     1381 gatccgtctt tgccgcatgc gacgctaata gaagcggcag actggaacgg gaagagttca
     1441 gagccctgtg caccgagctg cgtgtgagac ctgccgatgc agaagccgtg ttccaaagac
     1501 tggacgccga ccgggatgga gccatcacat tccaggagtt cgccagaggc ttcctgggaa
     1561 gcctgcgggg cggccggaga cgggactggg gccctctgga ccctgctcct gctgtttccg
     1621 aggccggacc tgagacccac gatagcgaag aagatgaagg agatgaggat gccgccgccg
     1681 ctctggccac atcttgcggc cccgccagcc ctgggagagc ctggcaggac ttccaggcta
     1741 gactgggcga cgaagccaag ttcatcccca gagaggagca ggtgagcacc ctgtaccaga
     1801 acatcaacct tgtcgagcct cggctgattc agccctacga gcacgtgatc aagaacttca
     1861 tcagggaaat cagactgcaa agcaccgaga tggaaaatct ggccatcgcc gtcaaacggg
     1921 cccaggataa ggccgccatg cagctgagcg agctggaaga agaaatggac cagagaatcc
     1981 aggccgccga acacaagaca agaaaggatg agaagagaaa ggccgaagag gccctcagcg
     2041 acctgaggcg gcagtacgag accgaagtgg gcgatctgca ggtgaccatc aagaaactac
     2101 gcaaactgga agagcagagc aagcgggtgt ctcagaaaga agacgtggcc gccctcaaga
     2161 aacagatcta cgacctgagc atggaaaacc agaaggtgaa gaaggacctg ctggaagctc
     2221 agaccaatat cgccttcctg caaagcgagc tggacgcgct gaagtccgac tacgccgacc
     2281 agagcctgaa caccgagcgg gacctggaga tcatccgggc ctacaccgag gacagaaact
     2341 cactggaaag acagatcgag atcctgcaga ccgccaacag aaagctgcac gactctaacg
     2401 acggactgcg gagcgccctt gagaacagct attctaagtt caacagaagc ctgcacatca
     2461 acaacatcag ccctggcaac accatcagta gaagcagccc taagttcatc ggccacagcc
     2521 cccagcctct gggctacgat aggtcaagca gaagctctta cgtggacgag gactgtgata
     2581 gcctggccct ctgtgacccc ctgcagcgga ccaactgcga agtggacagc ctgcctgaga
     2641 gctgcttcga cagtggactg agcacactga gggaccctaa cgagtacgac agcgaggtgg
     2701 aatacaagca ccagcgcggg ttccagagaa gccacggcgt gcaggagtct tttggaggcg
     2761 atgctagcga cacagacgtg ccagatatcc gcgatgagga gacatttggc ctggaggacg
     2821 tcgcctccgt gctggactgg aagccccagg gctccgtgag cgagggcagc atcgtgtcta
     2881 gcagcagaaa acctatctcc gccctgagcc cacagaccga cctggtggac gacaacgcca
     2941 aatctttcag cagccagaaa gcttataaga tcgtgctggc tggcgacgcc gctgtgggca
     3001 agtccagctt cctcatgaga ctgtgtaaaa atgagtttcg ggaaaacatt agcgccaccc
     3061 tgggcgtgga tttccaaatg aagaccctga tcgtagacgg cgagagaacc gtgctgcaac
     3121 tgtgggacac cgccggccag gagcgattta gatccattgc caagtcctac ttcagaaagg
     3181 ccgatggcgt gctgctgctg tacgacgtta catgcgagaa gtcttttctg aatatcaggg
     3241 agtgggtgga tatgatcgag gatgctgccc acgagacagt gcctatcatg ctggtgggaa
     3301 acaaggctga catccgggac accgccgcta ccgagggcca gaagtgcgtg cccggacatt
     3361 tcggcgagaa actggccatg acctacggcg ccctgttctg cgagactagc gccaaggacg
     3421 gctctaatat cgtggaagcc gtgctgcacc tggctcgaga agtgaagaag agaacagata
     3481 aggacgacag cagatctatc accaacctga ccggcacaaa cagcaagaag tctccacaga
     3541 tgaagaactg ttgcaacggc tgagaattct gcagtcgacg gtaccgcggg cccgggatcc
     3601 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3661 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3721 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3781 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3841 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3901 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3961 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4021 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4081 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4141 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4201 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4261 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     4321 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4381 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4441 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4501 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4561 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4621 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4681 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4741 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4801 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4861 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4921 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4981 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     5041 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5101 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5161 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5221 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5281 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5341 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5401 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5461 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5521 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5581 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5641 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5701 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5761 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5821 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5881 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5941 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     6001 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     6061 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     6121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6901 cctgattctg tggataaccg tattaccgcc atgcat
//
