LOCUS       sequence-486858-        5896 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Express Rab7a-HiBiT Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5896)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5896)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Rab7a-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        129..233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(234..302,303..1094)
                     /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(452..471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1252..1840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1332..1351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1956..2239
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        2430..2809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2810..3013
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2964..2984
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          3149..3281
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3264..3283
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     3325..3344
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3325..3343
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4134..4152)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4189..4310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4200..4219)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4254..4273
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4555..4912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4555..4746
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4578..4598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4763..4898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4963..5757
                     /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(5017..5036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5627..5646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5821..5869
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc
       61 gtttctgcgg actggctttc tacgtaatgg tttcttagac gtcaggtggc acttttcggg
      121 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      181 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      241 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      301 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      361 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      421 gttttccaat gatgagcact ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg
      481 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      541 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      601 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaact atcggaggac
      661 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      721 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      781 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      841 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      901 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      961 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1021 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1081 ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     1141 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1201 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1261 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1321 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1381 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1441 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1501 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1561 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1621 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1681 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1741 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1801 ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc
     1861 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1921 atcaaaaagg atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg
     1981 atggcccttc gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg
     2041 ggtaatgtgg cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc
     2101 gcgattgcgg cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc
     2161 gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt
     2221 ctgccctcgt agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga
     2281 ggatcgagat cttcaatatt ggccattagc catattattc attggttata tagcataaat
     2341 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
     2401 tggctcatgt ccaatatgac cgccatgttg gcattgatta ttgactagtt attaatagta
     2461 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2521 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2581 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2641 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     2701 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     2761 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2821 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2881 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2941 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3001 tataagcaga gctggtttag tgaaccgtca gatcactaga agctttattg cggtagttta
     3061 tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga acttaagctg
     3121 cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg
     3181 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     3241 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa
     3301 ttacagctct taaggctaga gtattaatac gactcactat agggctagcg atcgccatga
     3361 cctctaggaa gaaagtgttg ctgaaggtta tcatcctggg agattctgga gtcgggaaga
     3421 catcactcat gaaccagtat gtgaataaga aattcagcaa tcagtacaaa gccacaatag
     3481 gagctgactt tctgaccaag gaggtgatgg tggatgacag gctagtcaca atgcagatat
     3541 gggacacagc aggacaggaa cggttccagt ctctcggtgt ggccttctac agaggtgcag
     3601 actgctgcgt tctggtattt gatgtgactg cccccaacac attcaaaacc ctagatagct
     3661 ggagagatga gtttctcatc caggccagtc cccgagatcc tgaaaacttc ccatttgttg
     3721 tgttgggaaa caagattgac ctcgaaaaca gacaagtggc cacaaagcgg gcacaggcct
     3781 ggtgctacag caaaaacaac attccctact ttgagaccag tgccaaggag gccatcaacg
     3841 tggagcaggc gttccagacg attgcacgga atgcacttaa gcaggaaacg gaggtggagc
     3901 tgtacaacga atttcctgaa cctatcaaac tggacaagaa tgaccgggcc aaggcctcgg
     3961 cagaaagctg cagttgcgtt tctcaaggca gttcaggtgg ctcgagcggt gtgagcggct
     4021 ggcggctgtt caagaagatt agctaaggcc gcgactctag agtcgacctg caggcatgca
     4081 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     4141 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     4201 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4261 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4321 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     4381 aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc
     4441 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     4501 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     4561 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4621 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4681 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4741 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4801 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4861 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga
     4921 ttcttctgac acaacagtct cgaacttaag gctagagcca ccatgattga acaagatgga
     4981 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5041 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5101 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5161 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5221 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5281 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5341 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5401 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5461 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5521 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5581 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5641 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5701 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5761 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     5821 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     5881 ataaggatcc tctttg
//
