LOCUS       sequence-441132-        5441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a single-cell discriminating version of 
            mVenus-Q69M (ME) fluorescent protein. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5441)
  AUTHORS   Szelenyi ER, Navarrete JS, Murry AD, Zhang Y, Girven KS, Kuo L, 
            Cline MM, Bernstein MX, Burdyniuk M, Bowler B, Goodwin NL, Juarez B,
            Zweifel LS, Golden SA
  TITLE     An arginine-rich nuclear localization signal (ArgiNLS) strategy for 
            streamlined image segmentation of single cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Aug 6;121(32):e2320250121. doi: 
            10.1073/pnas.2320250121. Epub 2024 Jul 29.
  PUBMED    39074275
REFERENCE   2  (bases 1 to 5441)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-08-6"; volume: "121(32):e2320250121. 
            doi"; pages: " 10.1073/pnas.2320250121. Epub 2024 Jul 29"
COMMENT     Sequence Label: pAAV-EF1a-ArgiNLS-mVenus-Q69M(ME)
FEATURES             Location/Qualifiers
     source          1..5441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(1541..1569)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2164..2184
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2303..2323)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2705..2845
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2705..2834
                     /label=AAV2 ITR
     rep_origin      2920..3375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3007..3026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3217..3238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3392..3411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3511..3533
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3571..3589)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3657..3761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3762..3830,3831..4622)
                     /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(3980..3999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4793..5380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5281..5300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccgtacc ggatcctcta gagccaccat
     1441 gaaaagggtg gctccccaga agcagatgag catagcaaag aaaaagaagg tgagaaggcg
     1501 gcggcgccgt cgaagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     1561 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
     1621 aacatacgga aaacttaccc ttaaattgat ttgcactact ggaaaactac ctgttccatg
     1681 gccaacactt gtcactactt tgggttatgg tctgatgtgc tttgcgagat acccagatca
     1741 tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     1801 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
     1861 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     1921 tggacacaaa ttggaataca actataactc acacaatgta tacatcacgg cagacaaaca
     1981 aaagaatgga atcaaagcga acttcaaaat tagacacaac attgaagatg gaggtgttca
     2041 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     2101 caaccattac ctgtcctacc aatctaagct ttcgaaagat cccaacgaaa agagagacca
     2161 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     2221 caaataagat atcgaattcc gctcgagata atcaacctct ggattacaaa atttgtgaaa
     2281 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2341 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2401 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2461 gggctcggct gttgggcact gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta
     2521 ttgctttatt tgtaaccatc tagctttatt tgtgaaattt gtgatgctat tgctttattt
     2581 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2641 caggttcagg gggagatgtg ggaggttttt taaagcggcc gccacgtgcg gaccgagcgg
     2701 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2761 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2821 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     2881 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2941 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3001 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3061 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3121 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3181 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3241 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3301 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3361 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3421 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3481 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3541 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3601 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3661 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3721 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3781 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3841 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3901 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3961 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4021 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4081 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4141 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4201 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4261 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4321 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4381 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4441 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4501 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4561 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4621 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4681 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4741 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4801 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4861 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4921 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4981 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5041 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5101 ggtcgggctg aacgggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5161 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5221 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5281 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5341 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5401 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
