LOCUS       sequence-425981-        6752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RAB7A. N-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6752)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 6752)
  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: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pHTN-HaloTag_RAB7A:1-207
FEATURES             Location/Qualifiers
     source          1..6752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(2703..2721)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2753..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2764..2783)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2818..2837
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3119..3476
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3119..3310
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3142..3162)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3327..3462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3527..4321
                     /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(3581..3600)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4191..4210
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4385..4433
                     /note="synthetic polyadenylation signal"
     promoter        4589..4693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4694..4762,4763..5554)
                     /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     complement(4912..4931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5712..6300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5792..5811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6416..6699
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgcttccga attcatgacc tctaggaaga aagtgttgct
     2041 gaaggttatc atcctgggag attctggagt cgggaagaca tcactcatga accagtatgt
     2101 gaataagaaa ttcagcaatc agtacaaagc cacaatagga gctgactttc tgaccaagga
     2161 ggtgatggtg gatgacaggc tagtcacaat gcagatatgg gacacagcag gacaggaacg
     2221 gttccagtct ctcggtgtgg ccttctacag aggtgcagac tgctgcgttc tggtatttga
     2281 tgtgactgcc cccaacacat tcaaaaccct agatagctgg agagatgagt ttctcatcca
     2341 ggccagtccc cgagatcctg aaaacttccc atttgttgtg ttgggaaaca agattgacct
     2401 cgaaaacaga caagtggcca caaagcgggc acaggcctgg tgctacagca aaaacaacat
     2461 tccctacttt gagaccagtg ccaaggaggc catcaacgtg gagcaggcgt tccagacgat
     2521 tgcacggaat gcacttaagc aggaaacgga ggtggagctg tacaacgaat ttcctgaacc
     2581 tatcaaactg gacaagaatg accgggccaa ggcctcggca gaaagctgca gttgctgatg
     2641 atctagattt gggcccaatt cctgcaggat tttgcggccg cttgctgagt tggctgctgc
     2701 caccgctgag caataactag cataacccct tggccgcttc gagcagacat gataagatac
     2761 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2821 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2881 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     2941 aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat
     3001 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga
     3061 tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct
     3121 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3241 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3301 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3361 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3421 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3481 ttgattcttc tgacacaaca gtctcgaact taaggctaga gccaccatga ttgaacaaga
     3541 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3601 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3661 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3721 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3781 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3841 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3901 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3961 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4021 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4081 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4141 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4201 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4261 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4321 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
     4381 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4441 agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     4501 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg tttcttagac
     4561 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4621 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4681 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4741 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4801 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4861 gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc tgctatgtgg
     4921 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4981 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5041 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5101 tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5161 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5221 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5521 tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc
     5581 caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6121 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa
     6301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga
     6421 aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg aaaaatggca
     6481 gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat aggtattcat
     6541 acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt tgccattttt
     6601 acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt
     6661 atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca gtactccggc
     6721 cacgatgcgt ccggcgtaga ggatcgagat ct
//
