LOCUS       sequence-425980-        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RAB7A. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  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 6764)
  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: pHTC-HaloTag_RAB7A:1-207
FEATURES             Location/Qualifiers
     source          1..6764
                     /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"
     CDS             1718..1738
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1754..2638
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(2710..2728)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2765..2886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2776..2795)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2830..2849
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3131..3488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3131..3322
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3154..3174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3339..3474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3539..4333
                     /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(3593..3612)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4203..4222
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4397..4445
                     /note="synthetic polyadenylation signal"
     promoter        4601..4705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4706..4774,4775..5566)
                     /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(4924..4943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5724..6312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5804..5823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6428..6711
                     /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 gcgatcgctt ccgaattcac
     1081 catgacctct aggaagaaag tgttgctgaa ggttatcatc ctgggagatt ctggagtcgg
     1141 gaagacatca ctcatgaacc agtatgtgaa taagaaattc agcaatcagt acaaagccac
     1201 aataggagct gactttctga ccaaggaggt gatggtggat gacaggctag tcacaatgca
     1261 gatatgggac acagcaggac aggaacggtt ccagtctctc ggtgtggcct tctacagagg
     1321 tgcagactgc tgcgttctgg tatttgatgt gactgccccc aacacattca aaaccctaga
     1381 tagctggaga gatgagtttc tcatccaggc cagtccccga gatcctgaaa acttcccatt
     1441 tgttgtgttg ggaaacaaga ttgacctcga aaacagacaa gtggccacaa agcgggcaca
     1501 ggcctggtgc tacagcaaaa acaacattcc ctactttgag accagtgcca aggaggccat
     1561 caacgtggag caggcgttcc agacgattgc acggaatgca cttaagcagg aaacggaggt
     1621 ggagctgtac aacgaatttc ctgaacctat caaactggac aagaatgacc gggccaaggc
     1681 ctcggcagaa agctgcagtt gcctcgagcc aaccactgag gatctgtact ttcagagcga
     1741 taacgatgga tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct
     1801 gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct
     1861 gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac
     1921 ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg
     1981 ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga
     2041 agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg
     2101 caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg
     2161 ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg
     2221 ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt
     2281 ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga
     2341 ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat
     2401 cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct
     2461 gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag
     2521 cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa
     2581 cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccggtta
     2641 atagaattgg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     2701 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     2761 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2821 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2881 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     2941 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt
     3001 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3061 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     3121 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3421 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3481 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat
     3541 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3601 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3661 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3721 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3781 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3841 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3901 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3961 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4021 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4081 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4141 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4201 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4261 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4321 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4381 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4441 gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc
     4501 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtaat
     4561 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4621 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4681 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4741 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4801 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4861 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca ctttcaaagt
     4921 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4981 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5041 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5101 ggccaactta cttctgacaa ctatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5161 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5221 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5281 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5341 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5401 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5461 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5521 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaattcg aaatgaccga
     5581 ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5641 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5701 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5761 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5821 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5881 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5941 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6001 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6061 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6121 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6181 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6241 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6301 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6361 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6421 ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca
     6481 tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc
     6541 ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt
     6601 gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac
     6661 aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg
     6721 cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
