LOCUS       sequence-237598-        6846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses a nuclear-localized iRFP713 reporter 
            and improved Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6846)
  AUTHORS   Back S, Necarsulmer J, Whitaker LR, Coke LM, Koivula P, Heathward 
            EJ, Fortuno LV, Zhang Y, Yeh CG, Baldwin HA, Spencer MD, 
            Mejias-Aponte CA, Pickel J, Hoffman AF, Spivak CE, Lupica CR, 
            Underhill SM, Amara SG, Domanskyi A, Anttila JE, Airavaara M, Hope 
            BT, Hamra FK, Richie CT, Harvey BK
  TITLE     Neuron-Specific Genome Modification in the Adult Rat Brain Using 
            CRISPR-Cas9 Transgenic Rats.
  JOURNAL   Neuron. 2019 Feb 8. pii: S0896-6273(19)30062-5. doi: 
            10.1016/j.neuron.2019.01.035.
  PUBMED    30792150
REFERENCE   2  (bases 1 to 6846)
  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: "Neuron. 
            2019 Feb 8. pii: S0896-6273(19)30062-5. doi: 
            10.1016/j.neuron.2019.01.035."
COMMENT     Sequence Label: pOTTC1442 - pAAV CMV-IE Nuc-iRFP-2A-iCre
FEATURES             Location/Qualifiers
     source          1..6846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        205..508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        509..712
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     663..683
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             753..1700
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     CDS             1734..1754
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1758..1778
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1782..1802
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1833..1886
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1899..1919
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2973..3561
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3026..3046)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3444..3455)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3593..4069
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3792..3811)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4109..4249
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4109..4238
                     /label=AAV2 ITR
     rep_origin      4324..4779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4411..4430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4621..4642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4796..4815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4915..4937
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4975..4993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5061..5165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5166..5234,5235..6026)
                     /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(5384..5403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6197..6785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6686..6705
                     /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 gcgttagtta ttaatagtaa tcaattacgg ggtcattagt
      181 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      241 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      301 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      361 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      421 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      481 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      541 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      601 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      661 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      721 gaaccgtcag atccggtacc ggatccgcca ccatggctga aggatccgtc gccaggcagc
      781 ctgacctctt gacctgcgac gatgagccga tccatatccc cggtgccatc caaccgcatg
      841 gactgctgct cgccctcgcc gccgacatga cgatcgttgc cggcagcgac aaccttcccg
      901 aactcaccgg actggcgatc ggcgccctga tcggccgctc tgcggccgat gtcttcgact
      961 cggagacgca caaccgtctg acgatcgcct tggccgagcc cggggcggcc gtcggagcac
     1021 cgatcactgt cggcttcacg atgcgaaagg acgcaggctt catcggctcc tggcatcgcc
     1081 atgatcagct catcttcctc gagctcgagc ctccccagcg ggacgtcgcc gagccgcagg
     1141 cgttcttccg ccgcaccaac agcgccatcc gccgcctgca ggccgccgaa accttggaaa
     1201 gcgcctgcgc cgccgcggcg caagaggtgc ggaagattac cggcttcgat cgggtgatga
     1261 tctatcgctt cgcctccgac ttcagcggcg aagtgatcgc agaggatcgg tgcgccgagg
     1321 tcgagtcaaa actaggcctg cactatcctg cctcaaccgt gccggcgcag gcccgtcggc
     1381 tctataccat caacccggta cggatcattc ccgatatcaa ttatcggccg gtgccggtca
     1441 ccccagacct caatccggtc accgggcggc cgattgatct tagcttcgcc atcctgcgca
     1501 gcgtctcgcc cgtccatctg gaattcatgc gcaacatagg catgcacggc acgatgtcga
     1561 tctcgatttt gcgcggcgag cgactgtggg gattgatcgt ttgccatcac cgaacgccgt
     1621 actacgtcga tctcgatggc cgccaagcct gcgagctagt cgcccaggtt ctggcctggc
     1681 agatcggcgt gatggaagag ttgtacaagt ccggcctgcg cagccgcgcc gacccaaaaa
     1741 agaagcgcaa ggtagacccc aagaagaagc gcaaggtaga tccaaaaaag aagagaaagg
     1801 tactgtacaa gatgcattct agaggctccg gagagggcag gggaagtctt ctaacatgcg
     1861 gggacgtgga ggaaaatccc gggcccatcg atatggtgcc caagaagaag aggaaagtct
     1921 ccaacctgct gactgtgcac caaaacctgc ctgccctccc tgtggatgcc acctctgatg
     1981 aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct gaacacacct
     2041 ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg aacaacagga
     2101 aatggttccc tgctgaacct gaggatgtga gggactacct cctgtacctg caagccagag
     2161 gcctggctgt gaagaccatc caacagcacc tgggccagct caacatgctg cacaggagat
     2221 ctggcctgcc tcgcccttct gactccaatg ctgtgtccct ggtgatgagg agaatcagaa
     2281 aggagaatgt ggatgctggg gagagagcca agcaggccct ggcctttgaa cgcactgact
     2341 ttgaccaagt cagatccctg atggagaact ctgacagatg ccaggacatc aggaacctgg
     2401 ccttcctggg cattgcctac aacaccctgc tgcgcattgc cgaaattgcc agaatcagag
     2461 tgaaggacat ctcccgcacc gatggtggga gaatgctgat ccacattggc aggaccaaga
     2521 ccctggtgtc cacagctggt gtggagaagg ccctgtccct gggggttacc aagctggtgg
     2581 agagatggat ctctgtgtct ggtgtggctg atgaccccaa caactacctg ttctgccggg
     2641 tcagaaagaa tggtgtggct gccccttctg ccacctccca actgtccacc cgggccctgg
     2701 aagggatctt tgaggccacc caccgcctga tctatggtgc caaggatgac tctgggcaga
     2761 gatacctggc ctggtctggc cactctgcca gagtgggtgc tgccagggac atggccaggg
     2821 ctggtgtgtc catccctgaa atcatgcagg ctggtggctg gaccaatgtg aacattgtga
     2881 tgaactacat cagaaacctg gactctgaga ctggggccat ggtgaggctg ctcgaggatg
     2941 gggactgaga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     3001 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3061 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3121 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3181 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3241 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3301 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3361 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     3421 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3481 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3541 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     3601 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     3661 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     3721 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     3781 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     3841 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     3901 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     3961 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4021 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     4081 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4141 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4201 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4261 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4321 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4381 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4441 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4501 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4561 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4621 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4681 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4741 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4801 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4861 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4921 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4981 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5041 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5101 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5161 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5221 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5281 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5341 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5401 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5461 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5521 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5581 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5641 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5701 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5761 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5821 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5881 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5941 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6001 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6061 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6781 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6841 acatgt
//
