LOCUS       sequence-475079-        5896 bp    DNA     circular SYN 30-JUN-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)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5896)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Rab7a-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5896
                     /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(1842..1860)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1897..2018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1908..1927)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1962..1981
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2263..2620
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2263..2454
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2286..2306)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2471..2606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2671..3465
                     /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(2725..2744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3335..3354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3529..3577
                     /note="synthetic polyadenylation signal"
     promoter        3733..3837
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3838..3906,3907..4698)
                     /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(4056..4075)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4856..5444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4936..4955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5560..5843
                     /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 ggctagcgat cgccatgacc tctaggaaga
     1081 aagtgttgct gaaggttatc atcctgggag attctggagt cgggaagaca tcactcatga
     1141 accagtatgt gaataagaaa ttcagcaatc agtacaaagc cacaatagga gctgactttc
     1201 tgaccaagga ggtgatggtg gatgacaggc tagtcacaat gcagatatgg gacacagcag
     1261 gacaggaacg gttccagtct ctcggtgtgg ccttctacag aggtgcagac tgctgcgttc
     1321 tggtatttga tgtgactgcc cccaacacat tcaaaaccct agatagctgg agagatgagt
     1381 ttctcatcca ggccagtccc cgagatcctg aaaacttccc atttgttgtg ttgggaaaca
     1441 agattgacct cgaaaacaga caagtggcca caaagcgggc acaggcctgg tgctacagca
     1501 aaaacaacat tccctacttt gagaccagtg ccaaggaggc catcaacgtg gagcaggcgt
     1561 tccagacgat tgcacggaat gcacttaagc aggaaacgga ggtggagctg tacaacgaat
     1621 ttcctgaacc tatcaaactg gacaagaatg accgggccaa ggcctcggca gaaagctgca
     1681 gttgcgtttc tcaaggcagt tcaggtggct cgagcggtgt gagcggctgg cggctgttca
     1741 agaagattag ctaaggccgc gactctagag tcgacctgca ggcatgcaag ctgatccggc
     1801 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1861 ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     1921 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1981 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2041 ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca
     2101 aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     2161 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     2221 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     2281 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2341 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2401 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2461 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2521 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2581 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac
     2641 aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt
     2701 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2761 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2821 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2881 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2941 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3001 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3061 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3121 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3181 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3241 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3301 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3361 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3421 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3481 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct
     3541 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc
     3601 tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag
     3661 caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt
     3721 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3781 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3841 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3901 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3961 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4021 gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt
     4081 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4141 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4201 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aactatcgga
     4261 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4321 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4381 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4441 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4501 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4561 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4621 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4681 ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     4741 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4801 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4861 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4921 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4981 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5041 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5101 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5161 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5221 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5281 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5341 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5401 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     5461 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5521 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg
     5581 ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag
     5641 tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc
     5701 aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt
     5761 gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta
     5821 ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg
     5881 tagaggatcg agatct
//
