LOCUS       sequence-488900-        5922 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  Express HiBiT-Rab7a 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 5922)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5922)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-Rab7a Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5922
                     /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"
     primer_bind     complement(1868..1886)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1923..2044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1934..1953)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1988..2007
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2289..2646
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2289..2480
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2312..2332)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2497..2632
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2697..3491
                     /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(2751..2770)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3361..3380
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3555..3603
                     /note="synthetic polyadenylation signal"
     promoter        3759..3863
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3864..3932,3933..4724)
                     /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(4082..4101)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4882..5470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4962..4981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5586..5869
                     /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 ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgacct
     1141 ctaggaagaa agtgttgctg aaggttatca tcctgggaga ttctggagtc gggaagacat
     1201 cactcatgaa ccagtatgtg aataagaaat tcagcaatca gtacaaagcc acaataggag
     1261 ctgactttct gaccaaggag gtgatggtgg atgacaggct agtcacaatg cagatatggg
     1321 acacagcagg acaggaacgg ttccagtctc tcggtgtggc cttctacaga ggtgcagact
     1381 gctgcgttct ggtatttgat gtgactgccc ccaacacatt caaaacccta gatagctgga
     1441 gagatgagtt tctcatccag gccagtcccc gagatcctga aaacttccca tttgttgtgt
     1501 tgggaaacaa gattgacctc gaaaacagac aagtggccac aaagcgggca caggcctggt
     1561 gctacagcaa aaacaacatt ccctactttg agaccagtgc caaggaggcc atcaacgtgg
     1621 agcaggcgtt ccagacgatt gcacggaatg cacttaagca ggaaacggag gtggagctgt
     1681 acaacgaatt tcctgaacct atcaaactgg acaagaatga ccgggccaag gcctcggcag
     1741 aaagctgcag ttgcgtttaa acgaattcgg gctcggtacc cggggatcct ctagagtcga
     1801 cctgcaggca tgcaagctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1861 gctgccaccg ctgagcaata actagcataa ccccttgggg cggccgcttc gagcagacat
     1921 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1981 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2041 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     2101 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa
     2161 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2221 catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     2281 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     2341 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2401 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2461 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2521 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2581 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2641 tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taaggctaga gccaccatga
     2701 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2761 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2821 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     2881 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2941 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3001 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3061 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3121 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3181 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3241 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3301 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3361 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3421 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3481 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3541 atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     3601 gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc
     3661 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg
     3721 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3781 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3841 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3901 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3961 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4021 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc
     4081 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4141 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4201 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4261 ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4321 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4381 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4441 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4501 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4561 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4621 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4681 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc
     4741 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5281 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5461 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5581 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     5641 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     5701 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     5761 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     5821 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     5881 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
