LOCUS       pYSD101                 7288 bp ds-DNA     circular     12-NOV-2023
DEFINITION  .
COMMENT     Encodes the S. cerevisiae SUC2 (Invertase) protein (Part3b’) with
            the Aga2 signal peptide (Part3a’) assembled with pTEF1 promoter,
            tTDH1 terminator, LEU2 selectable marker and CEN/ARS yeast origin.
FEATURES             Location/Qualifiers
     misc_feature    1..194
                     /label="ConL1"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    5..10
                     /label="EcoRI Site"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    195..198
                     /label="BsaI"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    195..903
                     /label="pTEF1"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    195..903
                     /label="pYTK013 Part 2 pTEF1 [10-718]"
     promoter        199..898
                     /label="ScTEF1 Promoter"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    904..957
                     /label="AGA2 Signal Sequence"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             908..2578
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MQLLRCFSIFSVIASVLASMTNETSDRPLVHFTPNKGWMNDPNGLWYDEKDAKWHLYFQYNPNDTVWGTPLFWGHATSDDLTNWEDQPIAIAPKRNDSGAFSGSMVVDYNNTSGFFNDTIDPRQRCVAIWTYNTPESEEQYISYSLDGGYTFTEYQKNPVLAANSTQFRDPKVFWYEPSQKWIMTAAKSQDYKIEIYSSDDLKSWKLESAFANEGFLGYQYECPGLIEVPTEQDPSKSYWVMFISINPGAPAGGSFNQYFVGSFNGTHFEAFDNQSRVVDFGKDYYALQTFFNTDPTYGSALGIAWASNWEYSAFVPTNPWRSSMSLVRKFSLNTEYQANPETELINLKAEPILNISNAGPWSRFATNTTLTKANSYNVDLSNSTGTLEFELVYAVNTTQTISKSVFADLSLWFKGLEDPEEYLRMGFEVSASSFFLDRGNSKVKFVKENPYFTNRMSVNNQPFKSENDLSYYKVYGLLDQNILELYFNDGDVVSTNTYFMTTGNALGSVNMTTGVDNLFYIDGSDYKDDDDKDYKDDDDKDYKDDDDKGHHHHHH*"
     misc_feature    958..2478
                     /label="SUC2 (Invertase) Part 3b'"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    1992..1997
                     /label="Internal BsaI site"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     protein_bind    2479..2482
                     /label="BsaI"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    2479..2584
                     /label="3XFLAG6XHis"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    2585..2812
                     /label="tTDH1"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     terminator      2589..2812
                     /label="ScTDH1 Terminator"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     protein_bind    2813..2816
                     /label="BsaI"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    2813..2985
                     /label="ConR2"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     protein_bind    complement(2817..2820)
                     /label="BsmBI"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    2986..4906
                     /label="LEU2"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     promoter        2990..3637
                     /label="LEU2 Promoter"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     CDS             3638..4732
                     /label="LEU2"
                     /ApEinfo_revcolor="#7cff8c"
                     /ApEinfo_fwdcolor="#7cff8c"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHLIGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAFGLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTGDLGGSNSTTEVGDAVAEEVKKILA*"
     terminator      4733..4906
                     /label="AgLEU2 Terminator"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    4907..5418
                     /label="CEN6/ARS4"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     rep_origin      4911..5027
                     /label="CEN6"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     rep_origin      5028..5418
                     /label="ARS4"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    5419..7288
                     /label="AmpR-ColE1"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    5423..5430
                     /label="NotI site"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     terminator      complement(5431..5527)
                     /label="AmpR Terminator"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     CDS             complement(5528..6385)
                     /label="AmpR"
                     /ApEinfo_revcolor="#9f4240"
                     /ApEinfo_fwdcolor="#9f4240"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(6386..6516)
                     /label="AmpR Promoter"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     rep_origin      complement(6517..7280)
                     /label="ColE1"
                     /ApEinfo_revcolor="#7f7f7f"
                     /ApEinfo_fwdcolor="#7f7f7f"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    7281..7288
                     /label="NotI site"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
ORIGIN
        1 ccctgaattc gcatctagaa cggggtcatc acggctcatc atgcgccaaa caaatgtgtg
       61 caatacacgc tcggatgact gcatgatgac cgcactgact ggggacagca gatccaccta
      121 agcctgtgag agaagcagac acccgacaga tcaaggcagt tacgtctcac caaaccagat
      181 gtcaacacag ctacaacgcc ttgccaacag ggagttcttc agagacatgg aggctcaaaa
      241 cgaaattatt gacagcctag acatcaatag tcatacaaca gaaagcgacc acccaacttt
      301 ggctgataat agcgtataaa caatgcatac tttgtacgtt caaaatacaa tgcagtagat
      361 atatttatgc atattacata taatacatat cacataggaa gcaacaggcg cgttggactt
      421 ttaattttcg aggaccgcga atccttacat cacacccaat cccccacaag tgatccccca
      481 cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc
      541 cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc
      601 ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagacaccgc
      661 ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt
      721 gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
      781 taaacggtca tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
      841 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaag
      901 atcTATGatg cagttacttc gctgtttttc aatattttct gttattgctt cagttCtTGC
      961 TTCAATGACA AACGAAACTA GCGATAGACC TTTGGTCCAC TTCACACCCA ACAAGGGCTG
     1021 GATGAATGAC CCAAATGGGT TGTGGTACGA TGAAAAAGAT GCCAAATGGC ATCTGTACTT
     1081 TCAATACAAC CCAAATGACA CCGTATGGGG TACGCCATTG TTTTGGGGCC ATGCTACTTC
     1141 CGATGATTTG ACTAATTGGG AAGATCAACC CATTGCTATC GCTCCCAAGC GTAACGATTC
     1201 AGGTGCTTTC TCTGGCTCCA TGGTGGTTGA TTACAACAAC ACGAGTGGGT TTTTCAATGA
     1261 TACTATTGAT CCAAGACAAA GATGCGTTGC GATTTGGACT TATAACACTC CTGAAAGTGA
     1321 AGAGCAATAC ATTAGCTATT CTCTTGATGG TGGTTACACT TTTACTGAAT ACCAAAAGAA
     1381 CCCTGTTTTA GCTGCCAACT CCACTCAATT CAGAGATCCA AAGGTGTTCT GGTATGAACC
     1441 TTCTCAAAAA TGGATTATGA CGGCTGCCAA ATCACAAGAC TACAAAATTG AAATTTACTC
     1501 CTCTGATGAC TTGAAGTCCT GGAAGCTAGA ATCTGCATTT GCCAATGAAG GTTTCTTAGG
     1561 CTACCAATAC GAATGTCCAG GTTTGATTGA AGTCCCAACT GAGCAAGATC CTTCCAAATC
     1621 TTATTGGGTC ATGTTTATTT CTATCAACCC AGGTGCACCT GCTGGCGGTT CCTTCAACCA
     1681 ATATTTTGTT GGATCCTTCA ATGGTACTCA TTTTGAAGCG TTTGACAATC AATCTAGAGT
     1741 GGTAGATTTT GGTAAGGACT ACTATGCCTT GCAAACTTTC TTCAACACTG ACCCAACCTA
     1801 CGGTTCAGCA TTAGGTATTG CCTGGGCTTC AAACTGGGAG TACAGTGCCT TTGTCCCAAC
     1861 TAACCCATGG AGATCATCCA TGTCTTTGGT CCGCAAGTTT TCTTTGAACA CTGAATATCA
     1921 AGCTAATCCA GAGACTGAAT TGATCAATTT GAAAGCCGAA CCAATATTGA ACATTAGTAA
     1981 TGCTGGTCCC TGGTCgCGTT TTGCTACTAA CACAACTCTA ACTAAGGCCA ATTCTTACAA
     2041 TGTCGATTTG AGCAACTCGA CTGGTACCCT AGAGTTTGAG TTGGTTTACG CTGTTAACAC
     2101 CACACAAACC ATATCCAAAT CCGTCTTTGC CGACTTATCA CTTTGGTTCA AGGGTTTAGA
     2161 AGATCCTGAA GAATATTTGA GAATGGGTTT TGAAGTCAGT GCTTCTTCCT TCTTTTTGGA
     2221 CCGTGGTAAC TCTAAGGTCA AGTTTGTCAA GGAGAACCCA TATTTCACAA ACAGAATGTC
     2281 TGTCAACAAC CAACCATTCA AGTCTGAGAA CGACCTAAGT TACTATAAAG TGTACGGCCT
     2341 ACTGGATCAA AACATCTTGG AATTGTACTT CAACGATGGA GATGTGGTTT CTACAAATAC
     2401 CTACTTCATG ACCACCGGTA ACGCTCTAGG ATCTGTGAAC ATGACCACTG GTGTCGATAA
     2461 TTTGTTCTAC ATTGACggat ccgattacaa ggacgatgat gataaggact acaaagatga
     2521 tgatgacaaa gattataagg acgatgatga taaaggtcat catcatcatc atcactaact
     2581 cgagtggcat aaagcaatct tgatgaggat aatgattttt ttttgaatat acataaatac
     2641 taccgttttt ctgctagatt ttgtgatgac gtaaataagt acatattact ttttaagcca
     2701 agacaagatt aagcattaac tttacccttt tctttctaag tttcaatatt agttatcact
     2761 gtttaaaagt tatggcgaga acgtcggcgg ttaaaatata ttaccctgaa cggctggatg
     2821 tgagacgata aatcgcagcc aagtgagtga atagatgacg caccacggtc agacacggcc
     2881 acatcgtatc tcacaggagc aagcgcgata ggagcactca cacatagtac ggtgatccgc
     2941 tgactccttt gcccaaataa gacgtgagcc actagtgcac tgcagtacat cgaggagaac
     3001 ttctagtata tctacatacc taatattatt gccttattaa aaatggaatc ccaacaatta
     3061 catcaaaatc cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt
     3121 caaaaacgtt atatttatag gataattata ctctatttct caacaagtaa ttggttgttt
     3181 ggccgagcgg tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa
     3241 tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc
     3301 aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt
     3361 ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga
     3421 aaaaggaaag gtgagagcgc cggaaccggc ttttcatata gaatagagaa gcgttcatga
     3481 ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt
     3541 tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc
     3601 aatatatata tatatatttc aaggatatac cattctaatg tctgccccta agaagatcgt
     3661 cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta aggttcttaa
     3721 agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg
     3781 tgctgctatc gatgctacag gtgttccact tccagatgag gcgctggaag cctccaagaa
     3841 ggctgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggaaccg gtagtgttag
     3901 acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca acttaagacc
     3961 atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac aatttgctaa
     4021 aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg gtaagagaaa
     4081 ggaggacgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc cagaagtgca
     4141 aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat tgcctatttg
     4201 gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa ctgtggagga
     4261 aaccatcaag aacgaatttc ctacattgaa ggttcaacat caattgattg attctgccgc
     4321 catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca gcaacatgtt
     4381 tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc
     4441 tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca
     4501 cggttctgct ccagatttgc caaagaataa ggtcaaccct atcgccacta tcttgtctgc
     4561 tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc
     4621 agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac
     4681 caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aacctggagg
     4741 acccttctct ttagactatt ctactcttat gcacgtaaaa aattctagga aatatgtatt
     4801 aactaggagt aaaataaccg gctagtggca ttcatatagc cgtctgttta catctacatc
     4861 acacatttcg agtgtatatc tcgcaacgtt ggcgttaaat aggcaggagt atcacgtgct
     4921 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
     4981 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
     5041 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
     5101 ttgtttcatc ttgtctgtgt agaaaaccac acacgaaaat cctgtgattt tacattttac
     5161 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
     5221 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
     5281 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     5341 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     5401 acaggcaagc gatccgtccc gagcggccgc gattatcaaa aaggatcttc acctagatcc
     5461 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5521 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5581 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5641 gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa
     5701 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5761 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5821 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5881 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5941 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6001 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6061 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6121 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6181 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6241 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6301 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6361 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6421 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6481 gggttccgcg cacatttccc cgaaaagtgc cacctgtcat gaccaaaatc ccttaacgtg
     6541 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6601 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6661 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6721 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6781 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6841 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6901 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6961 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7021 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7081 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7141 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7201 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7261 ctgattctgt ggataaccgt gcggccgc
//