LOCUS       pHS1625                 7875 bp ds-DNA     circular     09-MAY-2025
DEFINITION  .
KEYWORDS    "marker:AmpR, AmpR"
FEATURES             Location/Qualifiers
     primer_bind     310..329
                     /label="T7"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_binding    329..351
                     /label="LacO"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     RBS             368..390
                     /label="RBS"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             397..900
                     /label="Translation 397-900"
                     /translation="MGSSHHHHSGHHHTGHHHHSGSHHHSSSGLVPRGSHMASWSHPQFEKGGGSGGGSGGSAWSHPQFEKGSAAGGEEDKKPAGGEGGGAHINLKVKGQDGNEVFFRIKRSTQLKKLMNAYCDRQSVDMTAIAFLFDGRRLRAEQTPDELEMEDGDEIDAMLHQTGGSSGS"
     CDS             397..2694
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MGSSHHHHSGHHHTGHHHHSGSHHHSSSGLVPRGSHMASWSHPQFEKGGGSGGGSGGSAWSHPQFEKGSAAGGEEDKKPAGGEGGGAHINLKVKGQDGNEVFFRIKRSTQLKKLMNAYCDRQSVDMTAIAFLFDGRRLRAEQTPDELEMEDGDEIDAMLHQTGGSSGSRYVYVLDVDGKPLMPTCRFGKVRRMLKSGQAKAVDTLPFTIQLTYKPRTRILQPVTLGQDPGRTNIGMAAVRFDGKELGRFHCITRNKEIPKLMADRMAARKASRRGERLARKRLARKLHTTAKHLNGRILPGCSEPIAVKDIINTESRFNNRILTKCKVCGKNTPLRRNVRELLLENIVRFLPLESELKETLKRTILEGQQGNINKLFRKLKFNQKDWPGKNLTDIAKNKLPGRLPFCKEHFAENEKFTTIEKSTFRLTPTATQLLRTHINLFRKLSGILPVTDVAVELNKFAFMQLDNPEMKKREIDFCHGPLCGTGGLEAAVKEQQDGKCLLCGKESIGHYHHIVPRSRRGSNTIANIAGLCPKCHELVHKDADTAESLTEMKTGLMKKYGGTSVLNQIIPKLVETLADLFPGHFHVTNGWNTKEFREKHHLEKDHDVDAYCIACSHLKPEETLVETEPFEILQFRKHNRAIIHHQTERTYKLDGVTVAKNRKKRMEQKTDSLEDWYVDMAKEHGKTQADAMRSRLTVIKSTRYYNTPGRMMPGTVFLYEGKRYVMTGQITNGKYYRAYGQEKRNFPAVKVRILTKNTGLVFVA*"
     CDS             409..474
                     /label="14xHIS"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             484..501
                     /label="thrombin site"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             514..537
                     /label="Twin-Strep-tag"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             538..573
                     /label="Twin-Strep-tag"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             574..597
                     /label="Twin-Strep-tag"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             598..888
                     /label="bdSUMO"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             889..900
                     /label="Linker"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    901..1359
                     /label="OrufIscB"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             901..2691
                     /label="Translation 901-2691"
                     /translation="RYVYVLDVDGKPLMPTCRFGKVRRMLKSGQAKAVDTLPFTIQLTYKPRTRILQPVTLGQDPGRTNIGMAAVRFDGKELGRFHCITRNKEIPKLMADRMAARKASRRGERLARKRLARKLHTTAKHLNGRILPGCSEPIAVKDIINTESRFNNRILTKCKVCGKNTPLRRNVRELLLENIVRFLPLESELKETLKRTILEGQQGNINKLFRKLKFNQKDWPGKNLTDIAKNKLPGRLPFCKEHFAENEKFTTIEKSTFRLTPTATQLLRTHINLFRKLSGILPVTDVAVELNKFAFMQLDNPEMKKREIDFCHGPLCGTGGLEAAVKEQQDGKCLLCGKESIGHYHHIVPRSRRGSNTIANIAGLCPKCHELVHKDADTAESLTEMKTGLMKKYGGTSVLNQIIPKLVETLADLFPGHFHVTNGWNTKEFREKHHLEKDHDVDAYCIACSHLKPEETLVETEPFEILQFRKHNRAIIHHQTERTYKLDGVTVAKNRKKRMEQKTDSLEDWYVDMAKEHGKTQADAMRSRLTVIKSTRYYNTPGRMMPGTVFLYEGKRYVMTGQITNGKYYRAYGQEKRNFPAVKVRILTKNTGLVFVA"
     misc_feature    1360..1674
                     /label="NbaCas9-1 REC"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    1675..2691
                     /label="OrufIscB"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    2695..900
                     /label="BB"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     terminator      2695..2742
                     /label="T7 terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        complement(2917..2922)
                     /label="tet promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        complement(2923..2939)
                     /label="tet promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        complement(2940..2945)
                     /label="tet promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        3059..3163
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             3164..3232
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             3233..4024
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             3362..4021
                     /label="AmpR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      4119..4801
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     rep_origin      4195..4783
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             complement(6716..7798)
                     /label="lacI"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     promoter        complement(7799..1)
                     /label="lacI promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
ORIGIN
        1 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
       61 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
      121 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
      181 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
      241 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
      301 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
      361 aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcCA TCACCATCAT
      421 TCAGGCCATC ACCATACCGG ACACCACCAT CATTCAGGCA GTCATCACCA TTCCagcagc
      481 ggcctggtgc cgcgcggcag ccatatggct agctggagcc atccgcagtt tgaaaaaggt
      541 ggtggtagcg gtggtggttc aggtggtagt gcatggtcac accctcagtt tgagaaaGGA
      601 TCTGCTGCGG GTGGCGAAGA AGATAAGAAA CCGGCAGGTG GCGAAGGTGG CGGTGCCCAT
      661 ATCAACCTGA AAGTGAAAGG TCAAGACGGC AACGAAGTCT TTTTCCGCAT CAAACGTTCT
      721 ACCCAGCTGA AAAAGCTGAT GAACGCATAC TGTGACCGTC AGTCTGTAGA CATGACCGCA
      781 ATTGCTTTCC TGTTTGATGG TCGTCGCCTG CGTGCGGAAC AGACCCCGGA TGAACTGGAG
      841 ATGGAAGATG GCGACGAAAT CGACGCAATG CTGCACCAGA CTGGTGGCTC AAGTGGATCC
      901 AGATACGTGT ACGTGCTGGA TGTGGACGGC AAGCCTCTGA TGCCTACATG CAGGTTCGGC
      961 AAGGTGCGGA GAATGCTGAA ATCTGGCCAA GCTAAGGCCG TGGACACCCT GCCTTTCACC
     1021 ATCCAGCTGA CCTACAAGCC CAGAACCAGA ATCCTGCAAC CTGTGACCCT GGGCCAGGAC
     1081 CCCGGCAGAA CCAATATCGG AATGGCCGCC GTGCGGTTCG ACGGCAAGGA ACTGGGAAGA
     1141 TTCCACTGCA TCACAAGAAA CAAGGAAATC CCTAAGCTGA TGGCCGACAG AATGGCAGCC
     1201 AGAAAGGCCT CCCGGAGAGG CGAGAGACTG GCCAGAAAGC GCCTGGCCCG GAAACTGCAC
     1261 ACCACCGCCA AGCACCTGAA TGGCAGAATC CTGCCAGGAT GTAGCGAGCC CATCGCCGTG
     1321 AAGGACATCA TCAACACAGA GAGCAGATTC AACAACAGAA TCCTGACCAA ATGCAAAGTG
     1381 TGCGGCAAGA ACACCCCTCT GAGAAGAAAC GTGCGGGAAC TCCTGCTGGA AAACATCGTG
     1441 CGGTTTCTGC CTCTGGAAAG CGAGCTGAAG GAAACACTCA AGAGAACCAT CCTGGAAGGC
     1501 CAGCAGGGCA ATATCAACAA GCTGTTCAGG AAACTTAAAT TCAATCAAAA GGATTGGCCT
     1561 GGCAAAAACC TGACCGATAT CGCTAAGAAT AAGCTGCCAG GCCGGCTGCC CTTCTGCAAG
     1621 GAGCACTTCG CCGAGAACGA GAAGTTCACC ACCATCGAGA AGAGCACCTT CCGGCTGACC
     1681 CCTACCGCCA CCCAGCTGCT GCGGACCCAC ATCAACCTGT TTAGAAAGCT GAGCGGCATC
     1741 CTGCCCGTGA CCGACGTGGC CGTCGAGCTG AACAAGTTCG CCTTTATGCA GCTGGACAAC
     1801 CCTGAGATGA AGAAGCGGGA AATCGACTTC TGCCACGGCC CACTGTGCGG CACCGGCGGC
     1861 CTGGAAGCCG CTGTTAAGGA GCAGCAGGAT GGAAAATGCC TGCTGTGTGG AAAAGAATCT
     1921 ATCGGCCACT ACCACCACAT CGTGCCTAGG TCCCGTAGAG GCAGCAACAC CATCGCTAAT
     1981 ATCGCCGGCC TTTGTCCTAA GTGCCACGAG CTCGTGCACA AGGACGCCGA TACAGCCGAG
     2041 TCTCTGACAG AGATGAAAAC CGGCCTGATG AAGAAGTATG GAGGCACCAG CGTGCTGAAC
     2101 CAGATCATCC CCAAGCTGGT GGAAACCCTG GCCGACCTGT TTCCTGGCCA CTTCCACGTG
     2161 ACAAACGGCT GGAACACCAA GGAGTTCAGA GAGAAACATC ATCTGGAAAA GGATCACGAC
     2221 GTGGATGCTT ACTGCATCGC CTGCAGCCAC CTGAAGCCTG AGGAAACCCT GGTGGAAACT
     2281 GAGCCTTTCG AGATCCTGCA GTTCAGAAAG CACAACAGAG CCATCATTCA CCACCAGACC
     2341 GAGCGGACGT ACAAGCTGGA CGGCGTGACA GTGGCTAAAA ATAGAAAGAA ACGGATGGAA
     2401 CAGAAAACCG ACAGCCTGGA AGATTGGTAC GTGGACATGG CCAAGGAACA CGGCAAAACA
     2461 CAGGCCGATG CCATGCGGAG CAGACTGACC GTGATCAAGA GCACACGGTA TTACAACACC
     2521 CCTGGCAGAA TGATGCCAGG CACAGTTTTC CTGTACGAGG GCAAGAGATA CGTGATGACC
     2581 GGCCAGATTA CAAACGGCAA GTACTACAGA GCCTACGGCC AGGAGAAGCG GAACTTCCCC
     2641 GCTGTCAAGG TGAGGATTCT GACAAAGAAC ACCGGACTAG TCTTTGTGGC Ctagctagca
     2701 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     2761 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     2821 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     2881 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     2941 gtcaaacatg agagattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     3001 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3541 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     3721 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4021 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4081 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4141 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4321 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4861 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4921 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     4981 ttttctcctt acgcatctgt gcggtatttc acaccgcata tatggtgcac tctcagtaca
     5041 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     5101 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5161 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5221 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     5281 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     5341 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     5401 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     5461 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     5521 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     5581 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     5641 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     5701 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     5761 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     5821 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc
     5881 caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt
     5941 tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc
     6001 ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg
     6061 gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat
     6121 ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg
     6181 tccagtgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg
     6241 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     6301 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     6361 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     6421 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     6481 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     6541 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     6601 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     6661 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     6721 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6781 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     6841 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     6901 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     6961 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     7021 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     7081 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     7141 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     7201 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     7261 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     7321 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     7381 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     7441 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     7501 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     7561 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     7621 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     7681 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     7741 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     7801 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     7861 gccattcgat ggtgt
//