LOCUS       CMV-TO-Che-Kir2.1-HA-tv 7298 bp ds-DNA     circular     02-SEP-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             complement(328..1188)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     CDS             complement(331..990)
                     /label="AmpR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     promoter        complement(1189..1287)
                     /label="bla promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        1551..2178
                     /label="CMV"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        1630..1933
                     /label="CMV enhancer"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     promoter        1934..2137
                     /label="CMV promoter"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     TATA_signal     2123..2129
                     /label="TATA box"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2139..2157
                     /label="TetO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2139..2178
                     /label="TetR binding sites"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2160..2178
                     /label="TetO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          2300..2332
                     /label="Che-Kir2.1 F"
                     /note="sequence: cttcgccaccgctagcatggtgagcaagggcga"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    2302..2309
                     /label="Kozak"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     CDS             2313..2315
                     /label="Translation 2313-2315"
     CDS             2316..3026
                     /label="mCherry"
                     /ApEinfo_revcolor=#ff0000
                     /ApEinfo_fwdcolor=#ff0000
     primer          complement(3006..3044)
                     /label="Che-Kir2.1 R"
                     /note="sequence: CACACTGCCCATggcgccagacttgtacagctcgtccat"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    3033..4340
                     /label="KCNJ2 (human)"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     CDS             3033..4340
                     /label="Translation 3033-4340"
     misc_feature    3378..3404
                     /label="HA tag"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    4179..4196
                     /label="ER export signal"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          4330..4364
                     /label="Kir2.1-tvmvs-F"
                     /note="sequence: AGTCGGAGATActcgagGGTGGGGGTTCAGAGACG"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             4341..4400
                     /label="Translation 4341-4400"
     CDS             4359..4379
                     /label="TVMVsite"
                     /ApEinfo_revcolor=#ff0000
                     /ApEinfo_fwdcolor=#ff0000
     CDS             4359..4382
                     /label="Translation 4359-4382"
     primer          complement(4380..4410)
                     /label="Kir.21-tvmvs-R"
                     /note="sequence: ggatgtgcttGGTACCAGAGCCGCCCCCCCC"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     CDS             4401..4403
                     /label="Translation 4401-4403"
     misc_feature    4401..4442
                     /label="ADRA2C retention"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             4404..4450
                     /label="Translation 4404-4450"
     CDS             4454..4483
                     /label="c-myc epitope"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     misc_feature    4483..4485
                     /label="New Stop"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     CDS             4496..4513
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     misc_feature    4514..4516
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor=#a187be
                     /ApEinfo_fwdcolor=#a187be
     polyA_signal    4535..4749
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     polyA_signal    4782..4805
                     /label="BGH pA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    5088..5135
                     /label="FRT"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_recomb     5102..5135
                     /label="FRT"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     gene            5143..6163
                     /label="Hygromycin"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     polyA_signal    6295..6425
                     /label="SV40 pA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     complement(6461..6481)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_binding    complement(6487..6509)
                     /label="LacO"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        complement(6513..6543)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     rep_origin      join(6808..7298,1..183)
                     /label="pUC ori"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     rep_origin      complement(join(6849..7298,1..233))
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
ORIGIN
        1 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
       61 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
      901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
      961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1321 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     1381 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     1441 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     1501 ctgcttaggg ttaggcgttt tgcgctgctt cgtacgggcc agatatacgc gttgacattg
     1561 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     1621 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     1681 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     1741 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     1801 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     1861 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     1921 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     1981 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2041 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     2101 taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct
     2161 ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag
     2221 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac
     2281 tctagcgttt aaacttaagc ttcgccaccg ctagcatggt gagcaagggc gaggaggata
     2341 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     2401 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2461 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     2521 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatTccc gactacttga
     2581 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     2641 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     2701 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2761 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     2821 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     2881 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     2941 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     3001 gcggcatgga cgagctgtac aagtctggcg ccATGGGCAG TGTGCGAACC AACCGCTACA
     3061 GCATCGTCTC TTCAGAAGAA GACGGTATGA AGTTGGCCAC CATGGCAGTT GCAAATGGCT
     3121 TTGGGAACGG GAAGAGTAAA GTCCACACCC GACAACAGTG CAGGAGCCGC TTTGTGAAGA
     3181 AAGATGGCCA CTGTAATGTT CAGTTCATCA ATGTGGGTGA GAAGGGGCAA CGGTATCTCG
     3241 CAGACATCTT CACCACGTGT GTGGACATTC GCTGGCGGTG GATGCTGGTT ATCTTCTGCC
     3301 TGGCTTTCGT CCTGTCATGG CTGTTTTTTG GCTGTGTGTT TTGGTTGATA GCTCTGCTCC
     3361 ATGGGGACCT GGATGCATAC CCATACGATG TTCCAGATTA CGCTTCCAAA GAGGGCAAGG
     3421 CTTGTGTGTC CGAGGTCAAC AGCTTCACGG CTGCCTTCCT CTTCTCCATT GAGACCCAGA
     3481 CAACCATAGG CTATGGTTTC AGATGTGTCA CGGATGAATG CCCAATTGCT GTTTTCATGG
     3541 TGGTGTTCCA GTCAATCGTG GGCTGCATCA TAGATGCTTT CATCATTGGC GCAGTCATGG
     3601 CCAAGATGGC AAAGCCAAAG AAGAGAAACG AGACTCTTGT CTTCAGTCAC AATGCCGTGA
     3661 TTGCCATGAG AGACGGCAAG CTGTGTTTGA TGTGGCGAGT GGGCAATCTT CGGAAAAGCC
     3721 ACTTGGTGGA AGCTCATGTT CGAGCACAGC TCCTCAAATC CAGAATTACT TCTGAAGGGG
     3781 AGTATATCCC TCTGGATCAA ATAGACATCA ATGTTGGGTT TGACAGTGGA ATAGATCGTA
     3841 TATTTCTGGT GTCCCCAATC ACTATAGTCC ATGAAATAGA TGAAGACAGT CCTTTATATG
     3901 ATTTGAGTAA ACAGGACATT GACAACGCAG ACTTTGAAAT CGTGGTCATA CTGGAAGGCA
     3961 TGGTGGAAGC CACTGCCATG ACGACACAGT GCCGTAGCTC TTATCTAGCA AATGAAATCC
     4021 TGTGGGGCCA CCGCTATGAG CCTGTGCTCT TTGAAGAGAA GCACTACTAC AAAGTGGACT
     4081 ATTCCAGGTT CCACAAAACT TACGAAGTCC CCAACACTCC CCTTTGTAGT GCCAGAGACT
     4141 TAGCAGAAAA GAAATATATC CTCTCAAATG CAAATTCATT TTGCTATGAA AATGAAGTTG
     4201 CCCTCACAAG CAAAGAGGAA GACGACAGTG AAAATGGAGT TCCAGAAAGC ACTAGTACGG
     4261 ACACGCCCCC TGACATAGAC CTTCACAACC AGGCAAGTGT ACCTCTAGAG CCCAGGCCCT
     4321 TACGGCGAGA GTCGGAGATA ctcgagGGTG GGGGTTCAGA GACGGTTAGA TTCCAGAGTG
     4381 GGGGGGGCGG CTCTGGTACC aagcacatcc tcttccgacg gaggagaagg ggcttcaggc
     4441 agTAGggccc GAAcaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca
     4501 tcataatcat cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     4561 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4621 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4681 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4741 tgctggggat gcggtgggct ctatggagat ctatcgatgt tctggggatg cggtgggctc
     4801 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4861 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4921 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4981 ctttccccgt caagctctaa atcgggggtc cctttagggt tccgatttag tgctttacgg
     5041 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc
     5101 cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc tgaactcacc
     5161 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     5221 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     5281 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     5341 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
     5401 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     5461 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
     5521 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     5581 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     5641 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     5701 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     5761 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     5821 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     5881 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     5941 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     6001 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     6061 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     6121 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt actacgagat
     6181 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6241 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6421 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7261 gctacagagt tcttgaagtg gtggcctaac tacggcta
//