LOCUS       pAAV-hSyn-Kir2.1Mut-2A- 6161 bp ds-DNA     circular     04-JUN-2023
DEFINITION  .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name:
            pAAV-hSyn-Kir2.1Mut-2A-mScarlet-KASH.gb
FEATURES             Location/Qualifiers
     repeat_region   13..153
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     repeat_region   24..153
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          156..191
                     /label="OR_hSyn1-CW3SL-6"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="sequence: ggccgcagatgctgcatgcgtatgagtgcaagtggg"
     promoter        172..627
                     /label="hSyn1(S) Promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          569..587
                     /label="MT_pAAV-EGFPCreERT2_FWD_Sequencing"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="sequence: cagcgctgcctcagtctgc"
     primer          569..587
                     /label="IS_hSyn-Seq5'"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="sequence: CAGCGCTGCCTCAGTCTGC"
     regulatory      653..659
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /regulatory_class="other"
     CDS             660..661
                     /label="Myc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation=""
     primer          660..679
                     /label="OR_Kir2.1-10_BspEI"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="sequence: ctgagagcgcagggtaccggatcctctagagtcgactgccaccatgggcagtgtgagaaccaac"
     misc_feature    660..2037
                     /label="Kir2.1 plasmid"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             662..1951
                     /label="Kir2.1"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="GSVRTNRYSIVSSEEDGMKLATMAVANGFGNGKSKVHTRQQCRSRFVKKDGHCNVQFINVGEKGQRYLADIFTTCVDIRWRWMLVIFCLAFVLSWLFFGCVFWLIALLHGDLDTSKVSKACVSEVNSFTAAFLFSIETQTTIAAAFRCVTDECPIAVFMVVFQSIVGCIIDAFIIGAVMAKMAKPKKRNETLVFSHNAVIAMRDGKLCLMWRVGNLRKSHLVGAHVRAQLLKSRITSEGEFIPLDQIDINVGFDSGIDRIFLVSPITIVHEIDEDSPLYDLSKQDIDNADFEIVVILEGMVEATAMTTQCRSSYLANEILWGHRYEPVLFEEKHYYKVDYSRFHKTYEVPNTPLCSARDLAEKKYILSNANSFCYENEVALTSKEEEEDSENGVPESTSTDSPPGIDLHNQASVPLEPRPLRRESEIGSG"
     primer          1395..1418
                     /label="DK_Kir2.1-11"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="sequence: ACCAGATAGACATCAATGTTGGTT"
     CDS             1952..2005
                     /label="T2A"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides."
                     /product="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      2015..2024
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /regulatory_class="other"
     primer          complement(2018..2037)
                     /label="OR_Kir2.1-6"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="sequence: tgaactccttgatcactgcctcgcccttgctcaccatggt"
     CDS             2021..2983
                     /label="mScarlet-KASH"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHSTGGMDELYKSGLRSREEEEETDSRMPHLDSPGSSQPRRSFLSRVIRAALPLQLLLLLLLLLACLLPASEDDYSCTQANNFARSFYPMLRYTNGPPPT*"
     primer          2038..2057
                     /label="OR_Kir2.1-7"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="sequence: tggtgagcaagggcgaggcagtgatcaaggagttca"
     CDS             2038..2716
                     /label="mScarlet"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="GSDQGVHAVQGAHGGLHERPRVRDRGRGRGPPLRGHPDRQAEGDQGWPPALLLGHPVPSVHVRLQGLHQAPRRHPRLL*AVLPRGLQVGARDELRGRRRRDRDPGHLPGGRHPDLQGEAPRHQLPS*RPRNAEEDNGLGSVHRAVVPRGRRAEGRH*DGPAPEGRRTLPGGLQDHLQGQEARADARRLQRRPQVGHHLPQRGLHRGGTVRTLRGPPLHRRHGRAVQ"
     misc_feature    2038..2980
                     /label="mScarlet INSERT"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(2328..2347)
                     /label="DK_Chronos-6"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="sequence: GGTCACGGTCACGGCGCCGC"
     primer          complement(2562..2580)
                     /label="DK_TVA-4"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="sequence: ggcttcttggccttgtagg"
     CDS             2717..2734
                     /label="linker_Kash"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="SGLRSR"
     CDS             2735..2980
                     /label="Kash"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="EEEEETDSRMPHLDSPGSSQPRRSFLSRVIRAALPLQLLLLLLLLLACLLPASEDDYSCTQANNFARSFYPMLRYTNGPPPT"
     primer          complement(2948..2966)
                     /label="OR_Kir2.1-8"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="sequence: aaattttgtaatccagaggttgattatcgcttgatatcgaattcttaggtgggaggtggcccgttggtgtaccgcagcat"
     misc_feature    3000..3250
                     /label="WPRE trunc. (+-)"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(3056..3076)
                     /label="WPRE-R"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          complement(3070..3087)
                     /label="MT_pAAV-EGFPCreERT2_REV_Sequencing"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="sequence: cagcgtatccacatagcg"
     primer          complement(3078..3097)
                     /label="IS_IMR9103"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="sequence: GGCATTAAAGCAGCGTATCC"
     misc_feature    3282..3363
                     /label="SV40 Poly(A) (+-)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     repeat_region   3436..3565
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     repeat_region   3436..3576
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     rep_origin      3651..4106
                     /label="f1 ori"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer_bind     complement(3738..3757)
                     /label="F1ori-R"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer_bind     3948..3969
                     /label="F1ori-F"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer_bind     complement(4123..4142)
                     /label="pRS-marker"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     4242..4264
                     /label="pGEX 3'"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(4302..4320)
                     /label="pBRforEco"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     promoter        4388..4492
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             4493..5353
                     /label="AmpR"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer_bind     complement(4711..4730)
                     /label="Amp-R"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     rep_origin      5524..6112
                     /label="ori"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer_bind     6013..6032
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
ORIGIN
        1 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
       61 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
      121 agggagtggc caactccatc actaggggtt cctgcggccg cagatgctgc atgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcagggt accggatcct ctagagtcga ctgccaccat
      661 gggcagtgtg agaaccaacc ggtacagcat tgtctcttcg gaggaagatg gcatgaagct
      721 ggccactatg gcagttgcca atggctttgg gaatggcaag agtaaagtcc atacccgaca
      781 acagtgcagg agccgctttg tgaagaaaga tggtcattgc aatgttcagt ttatcaacgt
      841 gggtgagaag ggacagaggt acctggcaga catctttact acctgtgtcg acatccgctg
      901 gaggtggatg ctggttatct tctgtcttgc cttcgtgctc tcctggctgt tctttggctg
      961 tgtgttttgg ttgatagccc tgctccatgg ggatctagat acttctaaag tgagcaaagc
     1021 atgcgtgtcg gaggttaaca gcttcacggc tgccttcctc ttctccatcg agacccagac
     1081 aaccattgcc gctgctttca ggtgtgtgac agacgaatgc ccaattgctg tcttcatggt
     1141 ggtattccag tcaattgtag gctgcatcat tgacgccttc atcattggtg cagtcatggc
     1201 gaagatggca aagccaaaga agagaaatga gactcttgtc ttcagtcaca atgctgtgat
     1261 tgccatgagg gatggcaaac tctgcttgat gtggagagtg ggtaaccttc gaaagagcca
     1321 ccttgtggga gctcatgtcc gggcacagct tctcaaatct aggatcactt cagaagggga
     1381 gtttatccct ttggaccaga tagacatcaa tgttggtttt gatagtggaa ttgaccgcat
     1441 atttctagtg tcccccatca ctatcgttca cgaaatagat gaagacagcc ctttatatga
     1501 cttgagtaag caggacattg acaatgcaga ctttgaaatt gttgtcatac tggaaggcat
     1561 ggtggaggcg actgccatga caactcaatg ccggagttcg tatctggcca atgaaattct
     1621 ctggggtcac cgctatgagc cagtgctctt tgaagagaaa cactactata aagtagacta
     1681 ttcaagattc cataagactt atgaagtacc taacaccccc ctttgtagtg ccagagactt
     1741 agcagagaag aaatacatcc tttcaaatgc aaattcattt tgctatgaaa atgaagttgc
     1801 cctaacaagc aaagaggaag aggaggatag tgagaacgga gtcccagaga gcacaagcac
     1861 agactcacct cctggcatag atctccacaa ccaggcaagc gtacctctag agcccaggcc
     1921 cttaaggcga gaatcggaga taggcagtgg agagggcaga ggaagtctgc taacatgcgg
     1981 tgacgtcgag gagaatcctg gcccagcgga tcccgccacc atggtgagca agggcgaggc
     2041 agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga
     2101 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2161 gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat
     2221 gtacggctcc agggccttca ccaagcaccc cgccgacatc cccgactact ataagcagtc
     2281 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac
     2341 cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg
     2401 caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc
     2461 caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg
     2521 cctgaaggac ggcggacgct acctggcgga cttcaagacc acctacaagg ccaagaagcc
     2581 cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga
     2641 ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca ccgGCGGCat
     2701 ggacgagctg tacaagtccg gactcagatc tcgagaggag gaggaggaga cagacagcag
     2761 gatgccccac ctcgacagcc ccggcagctc ccagccgaga cgctccttcc tctcaagggt
     2821 gatcagggca gcgctaccgt tgcagctgct tctgctgctg ctgctgctcc tggcctgcct
     2881 gctacctgcc tctgaagatg actacagctg cacccaggcc aacaactttg cccgatcctt
     2941 ctaccccatg ctgcggtaca ccaacgggcc acctcccacc taagaattcg atatcaagcg
     3001 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3061 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3121 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     3181 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3241 ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc atctagcttt
     3301 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3361 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     3421 ttttaaagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3481 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3541 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     3601 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3661 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3721 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3781 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3841 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3901 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3961 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     4021 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4081 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4141 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4201 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4261 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4321 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4381 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4501 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4561 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4621 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4681 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4741 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4801 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4861 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4921 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4981 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5041 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5101 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5161 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5221 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5281 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5341 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5401 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5461 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5521 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5581 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5641 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5701 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5761 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5821 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5881 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5941 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6001 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6061 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6121 caacgcggcc tttttacggt tcctggcctt ttgctggcct t
//