LOCUS       sequence-441122-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a single-cell discriminating version of 
            AausFP1 fluorescent protein. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  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 5447)
  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-AausFP1
FEATURES             Location/Qualifiers
     source          1..5447
                     /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(2309..2329)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2711..2851
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2711..2840
                     /label=AAV2 ITR
     rep_origin      2926..3381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3013..3032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3223..3244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3398..3417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3517..3539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3577..3595)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3663..3767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3768..3836,3837..4628)
                     /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(3986..4005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4799..5386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5287..5306
                     /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 cgaacgggat cccctgtggc caccatgtct tatggcgccc tgctgttccg
     1561 cgagaagatc ccctatgtgg tggaaatgga aggcgacgtg gaaggcatga agttcagcgt
     1621 cagaggcaaa ggacacggcg acgccaatac cggaaagatc gaggccagct tcatctgcac
     1681 aaccggcgaa ctgcctgtgc cttggagcag catcctgacc acagtgacat acggcgccca
     1741 gtgcttcgcc aagtatccca acgacatcaa ggactacccc aagagcgcca tgcctgaggg
     1801 ctacgtgcaa gagagaacca tcaccttcga gaacgacggc gtgtacaaga ccagagccga
     1861 agtgacctac gagaagggct ccgtgtacaa cagagtgacc ctgaacggca gcggcttcaa
     1921 gaaaggcggc aacatcctgg gcaagaaact tgagttcaac tacaacccgc actgcatcta
     1981 cgtgctgccc gacgtgcaga acaacggcat caagtgctac atcaacattg tgcacgacgt
     2041 gatcggcgga ggccagatca ttgctgctca ccagcagctg aatacccctc tcggaggtgg
     2101 ccccgtggac atccctcact atcaccacat tcaggcccac accatcctga gcaaggaccc
     2161 taaagaaacc cgggaccaca tgaacgtggt ggaagtgttc agagccatcg actgcaagac
     2221 cgcctacgcc taagatatcg aattccgctc gagataatca acctctggat tacaaaattt
     2281 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2341 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2401 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2461 ggacaggggc tcggctgttg ggcactgaca attccgtggt gtttatttgt gaaatttgtg
     2521 atgctattgc tttatttgta accatctagc tttatttgtg aaatttgtga tgctattgct
     2581 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2641 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcggccgcca cgtgcggacc
     2701 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2761 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2821 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     2881 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2941 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3001 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3061 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3121 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3181 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3241 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     3301 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3361 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     3421 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3481 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3541 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3601 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3661 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3721 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3781 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3841 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3901 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3961 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4021 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4081 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4141 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4201 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4261 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4321 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4381 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4441 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4501 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4561 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4621 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4681 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4741 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4921 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5101 cgcagcggtc gggctgaacg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5161 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5221 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5281 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5341 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5401 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
