LOCUS       sequence-472634-        6704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry- human RAB1A Q70L in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  AUTHORS   Tremel S, Ohashi Y, Morado DR, Bertram J, Perisic O, Brandt LTL, von
            Wrisberg MK, Chen ZA, Maslen SL, Kovtun O, Skehel M, Rappsilber J, 
            Lang K, Munro S, Briggs JAG, Williams RL
  TITLE     Structural basis for VPS34 kinase activation by Rab1 and Rab5 on 
            membranes.
  JOURNAL   Nat Commun. 2021 Mar 10;12(1):1564. doi: 10.1038/s41467-021-21695-2.
  PUBMED    33692360
REFERENCE   2  (bases 1 to 6704)
  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"; journalName: "Nat 
            Commun."; date: "2021-03-10"; volume: "12(1):1564. doi"; pages: " 
            10.1038/s41467-021-21695-2"
COMMENT     Sequence Label: pJB180
FEATURES             Location/Qualifiers
     source          1..6704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        236..615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      902..911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             908..1615
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1053..1071)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2296..2313)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2302..2526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2572..3000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2659..2678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2869..2890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3009..3029)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3014..3343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3194..3329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3256..3275
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3410..4204
                     /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(3464..3483)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4074..4093
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4378..4499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4415..4434)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4469..4488
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4548..4564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4548..4564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4561..4583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4572..4588
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4596..4602,4603..4620,4621..4626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4641..4662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4779..4796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4950..5538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5030..5049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5709..6500,6501..6569))
                     /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     6332..6351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6570..6674)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
      961 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1021 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     1081 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     1141 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaattg
     1201 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     1261 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     1321 ccccgtaatg cagtgtcgta ccatgggctg ggaggcctcc actgagcgga tgtaccccga
     1381 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     1441 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1501 caacgtcgac atcaagttgg acatcctttc ccacaacgag gactacacca tcgtggaaca
     1561 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagggtgc
     1621 gggagcagga tccatgtcca gcatgaatcc cgaatatgat tatttattca agttacttct
     1681 gattggcgac tcaggggttg gaaagtcttg ccttcttctt aggtttgcag atgatacata
     1741 tacagaaagc tacatcagca caattggtgt ggatttcaaa ataagaacta tagagttaga
     1801 cgggaaaaca atcaagcttc aaatatggga cacagcaggc ctggaaagat ttcgaacaat
     1861 cacctccagt tattacagag gagcccatgg catcatagtt gtgtatgatg tgacagatca
     1921 ggagtccttc aataatgtta aacagtggct gcaggaaata gatcgttatg ccagtgaaaa
     1981 tgtcaacaaa ttgttggtag ggaacaaatg tgatctgacc acaaagaaag tagtagacta
     2041 cacaacagcg aaggaatttg ctgattccct tggaattccg tttttggaaa ccagtgctaa
     2101 gaatgcaacg aatgtagaac agtctttcat gacgatggca gctgagatta aaaagcgaat
     2161 gggtcccgga gcaacagctg gtggtgctga gaagtccaat gttaaaattc agagcactcc
     2221 agtcaagcag tcaggtggag gttgctgcta agcggccgct cgagtctaga gggcccgttt
     2281 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2341 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2401 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2461 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2521 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2581 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2641 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2701 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2761 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2821 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2881 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2941 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3001 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3061 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3181 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3301 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3361 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3421 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3481 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3541 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3601 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3661 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3721 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3781 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3841 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3901 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     3961 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4021 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4081 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4141 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4201 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4261 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4321 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4381 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4441 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4501 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgt
//
