LOCUS       sequence-441120-        5815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent 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 5815)
  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 5815)
  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-CAG-FLEX-ArgiNLS-AausFP1
FEATURES             Location/Qualifiers
     source          1..5815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    325..446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(362..381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     416..435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    489..1077
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             595..606
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1004..1024
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1119..1152
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1223..1256
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2094..2127)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2190..2223)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(2258..2277)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2339..2361)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2640..2917)
                     /label=chicken beta-actin promoter
     repeat_region   3228..3357
                     /label=AAV2 ITR
     rep_origin      3429..3884
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3516..3535)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3726..3747
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3910..4014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4015..4083,4084..4875)
                     /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(4233..4252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5046..5634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5535..5554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cggcctcagt gagcgagcga
      121 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac
      181 ccgccatgct acttatctac gtagccatgc tctaggaaga tccttatcga ttttaccaca
      241 tttgtagagg ttttacttgc tttaaaaaac ctcccacatc tccccctgaa cctgaaacat
      301 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
      361 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      421 ttgtccaaac tcatcaatgt atcttatcat gtctgctcga agcggccgcc cgggtcgacg
      481 gtatcgatgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag
      541 acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag
      601 ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag
      661 cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag tgcccaacag
      721 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg
      781 gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt
      841 gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa
      901 ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga aagccatacg
      961 ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
     1021 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgattatc
     1081 gataagcttg atatcgaatt cgaacctctt cgagggacat aacttcgtat aatgtatgct
     1141 atacgaagtt atgggtcgat ggtgagatct ggactagagg gtcgatggtg atgcttggat
     1201 caaaatagga agaatcccaa tgataacttc gtataaagta tcctatacga agttatacta
     1261 gaattcggta ccgccaccat gaaaagggtg gctccccaga agcagatgag catagcaaag
     1321 aaaaagaagg tgagaaggcg gcggcgccgt cgaacgggat cccctgtggc caccatgtct
     1381 tatggcgccc tgctgttccg cgagaagatc ccctatgtgg tggaaatgga aggcgacgtg
     1441 gaaggcatga agttcagcgt cagaggcaaa ggacacggcg acgccaatac cggaaagatc
     1501 gaggccagct tcatctgcac aaccggcgaa ctgcctgtgc cttggagcag catcctgacc
     1561 acagtgacat acggcgccca gtgcttcgcc aagtatccca acgacatcaa ggactacccc
     1621 aagagcgcca tgcctgaggg ctacgtgcaa gagagaacca tcaccttcga gaacgacggc
     1681 gtgtacaaga ccagagccga agtgacctac gagaagggct ccgtgtacaa cagagtgacc
     1741 ctgaacggca gcggcttcaa gaaaggcggc aacatcctgg gcaagaaact tgagttcaac
     1801 tacaacccgc actgcatcta cgtgctgccc gacgtgcaga acaacggcat caagtgctac
     1861 atcaacattg tgcacgacgt gatcggcgga ggccagatca ttgctgctca ccagcagctg
     1921 aatacccctc tcggaggtgg ccccgtggac atccctcact atcaccacat tcaggcccac
     1981 accatcctga gcaaggaccc taaagaaacc cgggaccaca tgaacgtggt ggaagtgttc
     2041 agagccatcg actgcaagac cgcctacgcc taactcgagg ggggatccct agtataactt
     2101 cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa ttcgggtcga
     2161 tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt tatacgaagt
     2221 tatggggatc caattctttg ccaaaatgat gagacagcac aataaccagc acgttgccca
     2281 ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta gagccgccgg
     2341 tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag ccgtcccccc
     2401 gcggacagcc ccgagccgct ccccccgcac aaagggccct cccggagccc ctcaaggctt
     2461 tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt acagcccgga
     2521 ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca gagccggggc
     2581 gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc agcgactccc
     2641 gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc cataaaagga
     2701 aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgccy csccccsccc
     2761 csccccccsc cccccccccc cccccccggc gcgcgccccc cccccccccc cgcccccatc
     2821 gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga ggggggggag
     2881 atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc gatgactaat
     2941 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     3001 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     3061 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     3121 cctattggcg ttactatggg aacatacgtc attattgacg tcaatggtac tcttcctaga
     3181 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     3241 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3301 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3361 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3421 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3481 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3541 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3601 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3661 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3721 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3781 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3841 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     3901 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3961 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5641 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5701 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5761 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
