LOCUS       sequence-380122-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a bright monomeric red FP to label eucaryotic cell 
            nuclei. To be used in tissue clearing methods and other fluorescent 
            microscopy methods.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5797)
  AUTHORS   Wang Z, Romanski A, Mehra V, Wang Y, Brannigan M, Campbell BC, 
            Petsko GA, Tsoulfas P, Blackmore MG
  TITLE     Brain-wide analysis of the supraspinal connectome reveals anatomical
            correlates to functional recovery after spinal injury.
  JOURNAL   Elife. 2022 Jul 15;11. pii: 76254. doi: 10.7554/eLife.76254.
  PUBMED    35838234
REFERENCE   2  (bases 1 to 5797)
  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: "Elife."; 
            date: "2022-07-15"; volume: "11. pii: 76254. doi"; pages: " 
            10.7554/eLife.76254"
COMMENT     Sequence Label: pAAV2-CAG-mScarlet-I-Cdt1
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(95..114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      908..1496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1397..1416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1733..1862
                     /label=AAV2 ITR
     polyA_signal    2024..2145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2061..2080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2115..2134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2188..2776
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2294..2305
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2703..2723
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2857..3129)
                     /codon_start=1
                     /product="degron consisting of residues 30-120 of human 
                     Cdt1 (Zielke and Edgar, 2015)"
                     /label=Cdt1 (30-120)
                     /translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
                     DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
     regulatory      3858..3867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3918..3937)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3999..4021)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4300..4576)
                     /label=chicken beta-actin promoter
     repeat_region   4887..5016
                     /label=AAV2 ITR
     rep_origin      5088..5543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5175..5194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5385..5406
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5569..5673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5674..5742,5743..5797,1..737)
                     /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"
ORIGIN
        1 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
       61 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      121 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      181 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      241 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      301 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      361 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      421 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      481 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      541 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      601 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      661 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      721 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      781 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1021 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1561 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1621 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     1681 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctgcgcgc
     1741 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     1801 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     1861 cttgtagtta atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagat
     1921 ccttatcgat tttaccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacatct
     1981 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2041 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2101 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgctcgaa
     2161 gcggccgccc gggtcgacgg tatcgatgcg gggaggcggc ccaaagggag atccgactcg
     2221 tctgagggcg aaggcgaaga cgcggaagag gccgcagagc cggcagcagg ccgcgggaag
     2281 gaaggtccgc tggattgagg gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc
     2341 aggtggcaac acaggcgagc agccaaggaa aggacgatga tttccccgac aacaccacgg
     2401 aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga
     2461 gttccgccgt ggcaataggg agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg
     2521 tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc acgccacgtt
     2581 gcctgacaac gggccacaac tcctcataaa gagacagcaa ccaggattta tacaaggagg
     2641 agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc
     2701 cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt
     2761 aatccagagg ttgattatcg ataagcttga tatcgtcgac tgttgtacaa aagttctaaa
     2821 acgtcttctc gaagacccgt tttgagctag ctttcagatg gtgtcctggt cctgggcaga
     2881 tgtcagatga ggaggctgtc cagcggcagg ggtagacttc ttgatcttct ggcctggaga
     2941 tgggcatgca ggaatgtctg gagcctcagg tgtgctgggg ctagacactt cgtccacgga
     3001 cagtctcagc cttctccgag ctggtggtct ggcttgatct ctgccaggag cagcaggagg
     3061 tctggctctt ttccggctgc cagatgtagc agaggcaggg gctctcagag cgggtctagc
     3121 aggagatgga aattcgccgc tgccagcagc agatccagca gagcccttgt acagctcgtc
     3181 cattccgcct gtagagtgtc tgccttcgct ccgctcgtac tgttccacca cggtgtagtc
     3241 ctcgttgtgg ctggtgatgt ccagctttct gtccacgttg taggcgccag gcatctgcac
     3301 gggtttcttg gccttgtagg tggtcttgaa gtcggccagg tatctgccgc catccttcag
     3361 tctcagggcc atcttgatgt cgcccttcag cacgccatcc tcaggataca gtctctcggt
     3421 gctggcttcc cagcccattg tctttttctg catcacgggg ccatcaggtg gaaagttggt
     3481 gccccgcagc ttcactttgt agatcagggt gccatcttcc aggcttgtat cctgtgtcac
     3541 ggtcacggct ccgccgtcct cgaagttcat cactctctcc cacttgaagc cctcggggaa
     3601 gctctgcttg tagtagtcgg gaatatcggc agggtgcttg atgaaggccc ggctgccgta
     3661 cataaactga gggctcagaa tgtcccagct gaaaggcaga gggccgcctt tggtcacttt
     3721 cagcttggcg gtctgtgttc cctcataagg tctgccctcg ccttcgcctt cgatctcgaa
     3781 ctcgtggccg ttcatgctgc cttccatgtg caccttgaac cgcatgaact ctttgatcac
     3841 ggcttcgccc ttggaaacca tggtggcgaa ttcaagcttt ctagaggatc caattctttg
     3901 ccaaaatgat gagacagcac aataaccagc acgttgccca ggagctgtag gaaaaagaag
     3961 aaggcatgaa catggttagc agaggctcta gagccgccgg tcacacgcca gaagccgaac
     4021 cccgccctgc cccgtccccc ccgaaggcag ccgtcccccc gcggacagcc ccgagccgct
     4081 ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc acagaaaaga
     4141 aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc cgtcccgccc
     4201 gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc
     4261 ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt
     4321 tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct
     4381 ctgattggct gccgccgcac ctctccgcct csccccsccc cscccctcsc cccsccccsc
     4441 cccscctggc gcgcgccccc cccccccccc gcccccatcg ctgcacaaaa taattaaaaa
     4501 ataaataaat acaaaattgg gggtggggag gggggggaga tggggagagt gaagcagaac
     4561 gtggggctca cctcgaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     4621 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     4681 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     4741 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     4801 acatacgtca ttattgacgt caatggtact cttcctagag catggctacg tagataagta
     4861 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4921 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4981 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg
     5041 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     5101 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5161 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5221 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5281 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5341 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5401 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5461 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5521 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     5581 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5641 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5701 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5761 aaagtaaaag atgctgaaga tcagttgggt gcacgag
//
