LOCUS       sequence-328900-        5010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged affinity reagent.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5010)
  AUTHORS   O'Shaughnessy EC, Stone OJ, LaFosse PK, Azoitei ML, Tsygankov D, 
            Heddleston JM, Legant WR, Wittchen ES, Burridge K, Elston TC, Betzig
            E, Chew TL, Adalsteinsson D, Hahn KM
  TITLE     Software for lattice light-sheet imaging of FRET biosensors, 
            illustrated with a new Rap1 biosensor.
  JOURNAL   J Cell Biol. 2019 Sep 2;218(9):3153-3160. doi: 
            10.1083/jcb.201903019. Epub 2019 Aug 23.
  PUBMED    31444239
REFERENCE   2  (bases 1 to 5010)
  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: 
            "10.1083/jcb.201903019"; journalName: "J Cell Biol"; date: 
            "2019-09-2- 2"; volume: "218"; issue: "9"; pages: "3153-3160"
COMMENT     Sequence Label: RalGDS-YPet
FEATURES             Location/Qualifiers
     source          1..5010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(1308..1327)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1719..1748
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1912..2033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1949..1968)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2003..2022
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2040..2495)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2177..2198)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2389..2408
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2522..2626
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2628..2985
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2628..2819
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2651..2671)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2836..2971
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3020..3814
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3020..3814
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3074..3093)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3684..3703
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4002..4021)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4046..4093
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4422..5010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4911..4930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc atggactgct
      721 gtatcatccg cgtcagcctg gacgtggaca atggcaacat gtacaagagc atcctggtga
      781 ccagccaaga taaggctccg gctgtaatcc gcaaggccat ggacaaacac aacctggagg
      841 aggaggagcc ggaggactat gagctgctgc agattctctc agatgaccgg aagctgaaga
      901 tccctgaaaa cgccaacgtc ttctatgcca tgaactctac cgccaactat gactttgtcc
      961 tcaagaagcg gaccttctcc ggaggatccg gttcactcga gatggtgagc aagggcgaag
     1021 agctgttcac cggcgtggtg cccatcctgg tggagctgga cggcgacgtg aacggccaca
     1081 agttcagcgt gagcggcgag ggcgagggcg acgccaccta cggcaagctg accctgaagc
     1141 tgctgtgcac caccggcaag ctgcccgtgc cctggcccac cctggtgacc accctgggct
     1201 acggcgtgca gtgcttcgcc cggtaccccg accacatgaa gcagcacgac ttcttcaaga
     1261 gcgccatgcc cgagggctac gtgcaggagc ggaccatctt cttcaaggac gacggcaact
     1321 acaagacccg ggccgaggtg aagttcgagg gcgacaccct ggtgaaccgg atcgagctga
     1381 agggcatcga cttcaaggag gacggcaaca tcctgggcca caagctggag tacaactaca
     1441 acagccacaa cgtgtacatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1501 agatccggca caacatcgag gacggcggcg tgcagctggc cgaccactac cagcagaaca
     1561 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagagcg
     1621 ccctgttcaa ggaccccaac gagaagcggg accacatggt gctgctggag ttcctgaccg
     1681 ccgccggcat caccgagggc atggacgagc tgtacaagga gcagaagctg atctccgagg
     1741 aggacctgtg agaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga
     1801 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1861 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1921 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1981 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     2041 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2101 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2161 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2221 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2281 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2341 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2401 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2461 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2521 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2581 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2641 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2701 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2761 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2821 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2881 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2941 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3001 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3061 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3121 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3181 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3241 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3301 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3361 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3421 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3481 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3541 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3601 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3661 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3721 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3781 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3841 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3901 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3961 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4021 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4081 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4141 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4201 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4261 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4321 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4381 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4441 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4501 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4561 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4621 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4681 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4741 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4801 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4861 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4921 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4981 tgctcgtcag gggggcggag cctatggaaa
//
