LOCUS       sequence-481876-        6726 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  AAV version of PS Intein palsmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6726)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 6726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AAV-PS Intein
FEATURES             Location/Qualifiers
     source          1..6726
                     /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        164..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        468..671
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     622..642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(848..866)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1116..1135
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2023..2472
                     /codon_start=1
                     /product="""positive Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=pMag
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with slow switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLIQIMNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     CDS             2815..2868
                     /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             join(2869..2871,2872..2874,2875..3588)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2914..2935)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3175..3194)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3522..3543
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3658..3678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     3860..3879
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3989..4129
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3989..4118
                     /label=AAV2 ITR
     rep_origin      4204..4659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4291..4310)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4501..4522
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4676..4695)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4795..4817
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4855..4873)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4941..5045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5046..5114,5115..5906)
                     /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(5264..5283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6077..6665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6566..6585
                     /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 gcggccgaag atgctgcaca cttgtgggct agccgttaca taacttacgg
      181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      661 taagcagagc tggtttagtg aaccgtcaga tccgctagca ccatggtgag caagggcgag
      721 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
      781 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
      841 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
      901 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
      961 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     1021 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     1081 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     1141 ggctgggagg cctcctccga gcggatgtac tccggatact gtctggatct gaaaacccag
     1201 gttcagacac cgcagggtat gaaagaaatt tcaaatattc aggtgggtga tctggttctg
     1261 agcaatactg gttataatga agtgctgaat gtgtttccga aaagcaaaaa aaaaagctat
     1321 aaaatcaccc tggaagatgg caaagaaatc atttgtagcg aagaacacct gtttccgacc
     1381 cagactggtg aaatgaatat tagcggtggt ctgaaagaag gtatgtgcct gtatgttaaa
     1441 gaaatgatgc tgaaaaaaat cctgaaaatc gaggaactgg atgaacgcgg taccggcagc
     1501 ggaggtcata ccttgtatgc gccggggggt tatgacatca tgggttacct catacagatc
     1561 atgaataggc cgaacccaca agtggaactc ggacccgtcg atacctcctg cgctctcatt
     1621 ctgtgtgacc ttaagcagaa ggatacccct atcgtgtacg cctccgaggc atttctgtac
     1681 atgacagggt actcgaacgc cgaagtgctg ggacggaact gccgcttcct gcaaagcccg
     1741 gatggaatgg tgaagcctaa gtcaacccgc aaatacgtgg actccaacac tatcaacacc
     1801 atgcgcaagg ccattgaccg caatgctgag gtgcaagtgg aagtggtgaa cttcaagaag
     1861 aatggacagc gcttcgtcaa cttcctgact atgatccccg tgcgcgacga gaccggcgaa
     1921 tatcggtaca gcatggggtt tcagtgtgag acagagggcg gctcaggagg cagcggcggt
     1981 tctggaggtt ccggtggcgg ctcaggaggt agcggagggt ctcacactct ttacgcccct
     2041 ggaggatacg acattatggg atatttgatt cagattatga accgcccaaa ccctcaggtc
     2101 gaactggggc ctgtggacac gtcatgtgcc ctgatcctgt gcgatctgaa gcaaaaggac
     2161 actccgattg tctacgcctc ggaagccttc ttgtatatga ccggatacag caatgcagag
     2221 gtgctcggta ggaactgcag attcctgcag tcccccgacg ggatggtgaa accaaagtcc
     2281 actcgcaaat atgtggactc gaacacgatc aatacaatgc ggaaggccat cgaccggaac
     2341 gccgaggtcc aggtggaggt ggtcaacttt aagaagaacg gccagcggtt cgtgaacttt
     2401 ctcaccatga ttccggtccg ggatgaaacc ggagagtaca gatactccat gggattccag
     2461 tgcgaaaccg aagggtcagg atccgaactg attgatattg aagttagcgg taaccacctg
     2521 ttctatgcca atgatattct gacccataat agctcgagcc ccgaggacgg cgccctgaag
     2581 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     2641 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     2701 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     2761 ggccgccact ccaccggcgg catggacgag ctgtacaagg aattcggtag cggcgagggc
     2821 agaggaagtc tgctaacatg cggtgatgtc gaggagaatc ctggcccaat ggtgagcaag
     2881 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2941 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3001 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3061 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3121 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3181 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3241 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3301 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     3361 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     3421 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3481 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3541 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga attccgctcg
     3601 agataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3661 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3721 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     3781 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3841 ttccgtggtg tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3901 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3961 gatgtgggag gttttttaaa gcggccgcag gaacccctag tgatggagtt ggccactccc
     4021 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4081 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4141 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4201 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4261 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4321 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4381 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4441 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4501 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4561 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4621 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4681 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4741 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4801 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4861 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4921 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4981 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5041 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5101 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5161 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5221 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5281 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5341 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5401 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5461 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5521 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5581 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5641 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5701 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5761 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5821 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5881 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6721 acatgt
//
