LOCUS       pHS1514_FLAG-SV40_NLS-C 7054 bp ds-DNA     circular     07-APR-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     promoter        complement(850..948)
                     /label="bla promoter"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        1217..1804
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    1743..1761
                     /label="CMV For"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer          1754..1774
                     /label="CMV forward primer"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        1848..1864
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          1848..1867
                     /label="T7 primer"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    1867..1896
                     /label="HindIII GH"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1896..1901
                     /label="HindIII"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             1908..3581
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MDYKDDDDKMAGSPKKKRKVGSMYCAAVCFPQTKYQQRGTALKKPLVGLRKMGVEAAAWNTLKVTRDRPKLTFPDLITPQSKFLDNDLWLKYKVPIEQKEHVMYNLLCDNWVNVVYLSGKPDRISLVQTLKDAHCLQLAYSNPMDRFTVFRFLLALGYWCFANTNVEPEPDKPLPVSWIPWLEENKEYFELFGDGKRFFQADPSSRIRAITDLIHEIPTAHNLCHFKHVTDYIDGLCEACCIKGLLRLPVFTTVGGRGIGAGINNTPPFYLLWHANDLAGMLAQNWQPWDNMGIPAWLGSFQKESREVGLLAGMTWLPRKVYLHDPVPGQAACCSCGLPSEALVYSCSIEVEPVPKGLEWKDPHGVYTDQGKSLQSKIKLMSNDRYTFADRDWYSPLFSYLHAEGNSRQGKLWLVGFASDKAKSIDIWDKIIELEGTDTNDELLAQLANRATALNAMRKKPLRGDFKKSVGTPQIADIIPHAENRIAINAGKMTENRGYSWQDADTEYGELLTKVAYSLEPAQTVDARLKRGNFISRKPWPIIPESKTKPAEGDQNE*"
     misc_feature    1911..1934
                     /label="FLAG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1941..1967
                     /label="SV40"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    1947..1968
                     /label="gibson annealing"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    1974..3581
                     /label="Cas5-HNH Cas8"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             1974..3581
                     /label="Translation 1974-3581"
                     /translation="MYCAAVCFPQTKYQQRGTALKKPLVGLRKMGVEAAAWNTLKVTRDRPKLTFPDLITPQSKFLDNDLWLKYKVPIEQKEHVMYNLLCDNWVNVVYLSGKPDRISLVQTLKDAHCLQLAYSNPMDRFTVFRFLLALGYWCFANTNVEPEPDKPLPVSWIPWLEENKEYFELFGDGKRFFQADPSSRIRAITDLIHEIPTAHNLCHFKHVTDYIDGLCEACCIKGLLRLPVFTTVGGRGIGAGINNTPPFYLLWHANDLAGMLAQNWQPWDNMGIPAWLGSFQKESREVGLLAGMTWLPRKVYLHDPVPGQAACCSCGLPSEALVYSCSIEVEPVPKGLEWKDPHGVYTDQGKSLQSKIKLMSNDRYTFADRDWYSPLFSYLHAEGNSRQGKLWLVGFASDKAKSIDIWDKIIELEGTDTNDELLAQLANRATALNAMRKKPLRGDFKKSVGTPQIADIIPHAENRIAINAGKMTENRGYSWQDADTEYGELLTKVAYSLEPAQTVDARLKRGNFISRKPWPIIPESKTKPAEGDQNE*"
     polyA_site      3639..3863
                     /label="BGH pA"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     rep_origin      3909..4337
                     /label="f1 origin"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     promoter        4342..4711
                     /label="SV40 early promoter"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     CDS             4747..5541
                     /label="Neo(R)"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     polyA_signal    5715..5845
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     rep_origin      complement(6228..6898)
                     /label="pUC origin"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             complement(7043..849)
                     /label="Amp(R)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
ORIGIN
        1 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
       61 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      121 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      181 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      241 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      301 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      361 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      421 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      481 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      541 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      601 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      661 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      721 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      781 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      841 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      901 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      961 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     1021 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     1081 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     1141 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     1201 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     1261 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     1321 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     1381 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     1441 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     1501 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     1561 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     1621 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     1681 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     1741 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     1801 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     1861 tatagggaga cccaagctgg ctagcgttta aacttaagct tgccaccATG gactataagg
     1921 atgatgacga caagatggcc ggatcTCCAA AAAAGAAACG GAAGGTGGGA tctATGTATT
     1981 GCGCGGCGGT TTGCTTTCCG CAGACAAAAT ATCAGCAAAG GGGAACTGCT CTAAAGAAAC
     2041 CGCTCGTCGG TCTGCGTAAG ATGGGTGTGG AAGCGGCTGC ATGGAACACC CTGAAGGTTA
     2101 CTAGAGATCG CCCTAAGTTG ACCTTTCCGG ATCTGATCAC TCCGCAAAGC AAATTCCTGG
     2161 ATAACGACTT ATGGCTGAAG TACAAAGTGC CAATTGAGCA GAAAGAACAC GTCATGTATA
     2221 ACCTGTTGTG CGATAACTGG GTTAATGTTG TTTACTTGAG CGGTAAACCG GACCGTATTT
     2281 CCTTAGTCCA AACCCTGAAG GACGCCCATT GTCTGCAATT GGCGTATTCT AACCCGATGG
     2341 ATCGTTTCAC CGTGTTCCGC TTCCTGTTGG CGTTGGGTTA CTGGTGTTTC GCGAACACGA
     2401 ACGTCGAGCC GGAGCCCGAC AAACCGCTGC CGGTGAGCTG GATTCCGTGG CTGGAGGAAA
     2461 ATAAAGAGTA CTTCGAGCTG TTCGGTGATG GTAAACGTTT CTTCCAAGCT GACCCGAGCA
     2521 GCCGTATTCG TGCGATTACT GACCTGATCC ATGAAATTCC GACCGCTCAC AACCTGTGCC
     2581 ATTTTAAGCA CGTGACCGAC TATATCGACG GCCTGTGTGA AGCCTGCTGC ATAAAAGGTC
     2641 TGTTGCGTCT GCCGGTATTT ACCACCGTTG GCGGTCGTGG CATTGGCGCA GGTATCAATA
     2701 ACACGCCACC GTTTTATCTG CTGTGGCATG CGAACGACTT AGCGGGCATG TTGGCGCAGA
     2761 ATTGGCAGCC TTGGGACAAC ATGGGCATCC CGGCGTGGCT GGGCTCTTTT CAGAAAGAGT
     2821 CGCGGGAAGT TGGCCTGCTT GCGGGTATGA CCTGGCTTCC GCGTAAAGTT TACCTGCACG
     2881 ATCCGGTTCC GGGTCAGGCT GCGTGCTGCA GCTGTGGTTT GCCGAGTGAA GCTCTGGTTT
     2941 ACTCATGCTC CATTGAGGTG GAACCAGTGC CAAAGGGCTT GGAGTGGAAG GACCCGCACG
     3001 GCGTTTACAC CGACCAGGGC AAATCCCTGC AGTCCAAAAT CAAGCTGATG AGCAATGACC
     3061 GCTATACCTT CGCGGACCGT GACTGGTATT CTCCGTTGTT CTCGTATCTT CACGCCGAAG
     3121 GTAATAGCCG CCAGGGTAAG CTGTGGCTGG TGGGTTTTGC GAGCGATAAA GCAAAATCTA
     3181 TCGACATCTG GGATAAGATC ATCGAGCTGG AGGGCACCGA CACCAATGAC GAACTGTTGG
     3241 CGCAACTCGC CAATCGTGCA ACCGCGCTGA ATGCAATGCG TAAAAAGCCG CTACGCGGTG
     3301 ATTTTAAAAA GAGCGTTGGC ACCCCGCAGA TTGCGGATAT CATCCCGCAT GCGGAAAACC
     3361 GCATCGCCAT TAACGCAGGC AAAATGACGG AAAACCGTGG TTACTCTTGG CAAGATGCCG
     3421 ATACGGAGTA CGGAGAACTG CTGACCAAGG TGGCATACTC GCTGGAGCCG GCGCAAACGG
     3481 TGGATGCTCG TCTGAAGCGC GGTAACTTTA TCAGCCGTAA GCCGTGGCCG ATTATCCCCG
     3541 AGAGCAAAAC AAAACCCGCA GAGGGGGACC AGAATGAATA GGGAtctTGA ggccgctcga
     3601 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3661 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3721 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3781 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3841 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3901 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3961 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4021 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4081 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4141 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4201 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4261 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4321 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4441 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4501 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4561 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4621 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4681 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4741 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4801 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4861 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4921 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4981 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5041 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5101 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5161 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5221 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5281 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5341 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5401 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5461 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5521 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5581 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5641 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5701 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5821 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5881 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5941 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6001 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6061 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6841 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6901 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6961 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7021 atgagtaaac ttggtctgac agttaccaat gctt
//