LOCUS       sequence-338220-        5333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Rab7a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5333)
  AUTHORS   Wu H, Voeltz GK
  TITLE     Reticulon-3 Promotes Endosome Maturation at ER Membrane Contact 
            Sites.
  JOURNAL   Dev Cell. 2021 Jan 11;56(1):52-66.e7. doi: 
            10.1016/j.devcel.2020.12.014.
  PUBMED    33434526
REFERENCE   2  (bases 1 to 5333)
  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"; doi: 
            "10.1016/j.devcel.2020.12"; journalName: "Dev Cell"; date: 
            "2021-01-11- 11"; volume: "56"; issue: "1"; pages: "52-66.e7"
COMMENT     Sequence Label: mScarlet-Rab7
FEATURES             Location/Qualifiers
     source          1..5333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             650..1345
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(783..801)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1051..1070
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2158..2279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2195..2214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2249..2268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2286..2741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2423..2444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2635..2654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2768..2872
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2874..3231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2874..3065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2897..2917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3082..3217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3266..4060
                     /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             3266..4060
                     /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(3320..3339)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3930..3949
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4248..4267)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4292..4339
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4668..5256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5157..5176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
      661 gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa
      721 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      781 cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc
      841 tcagttcatg tacggctcca gggccttcac caagcacccc gccgacatcc ccgactacta
      901 taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      961 cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa
     1021 gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg
     1081 ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat
     1141 ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc
     1201 caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc
     1261 ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac
     1321 cggcggcatg gacgagctgt acaagtccgg actcagatct cgagctatga cctctaggaa
     1381 gaaagtgttg ctgaaggtta tcatcctggg agattctgga gtcgggaaga catcactcat
     1441 gaaccagtat gtgaataaga aattcagcaa tcagtacaaa gccacaatag gagctgactt
     1501 tctgaccaag gaggtgatgg tggatgacag gctagtcaca atgcagatat gggacacagc
     1561 aggacaggaa cggttccagt ctctcggtgt ggccttctac agaggtgcag actgctgcgt
     1621 tctggtattt gatgtgactg cccccaacac attcaaaacc ctagatagct ggagagatga
     1681 gtttctcatc caggccagtc cccgagatcc tgaaaacttc ccatttgttg tgttgggaaa
     1741 caagattgac ctcgaaaaca gacaagtggc cacaaagcgg gcacaggcct ggtgctacag
     1801 caaaaacaac attccctact ttgagaccag tgccaaggag gccatcaacg tggagcaggc
     1861 gttccagacg attgcacgga atgcacttaa gcaggaaacg gaggtggagc tgtacaacga
     1921 atttcctgaa cctatcaaac tggacaagaa tgaccgggcc aaggcctcgg cagaaagctg
     1981 cagttgctga aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga
     2041 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2101 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2161 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2221 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2281 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2341 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2401 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2461 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2521 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2581 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2641 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2701 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     2761 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2821 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     2881 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3061 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3121 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3181 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3241 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3301 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3361 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3421 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3481 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3541 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3601 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3661 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3721 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3781 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3841 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3901 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3961 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4021 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4081 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4141 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4201 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4261 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4321 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4381 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4441 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4501 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4561 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4621 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4681 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4741 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4801 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4861 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4921 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4981 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5041 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5101 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5161 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5221 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5281 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atc
//
