LOCUS       pYSD140                 6427 bp ds-DNA     circular     12-NOV-2023
DEFINITION  .
COMMENT     Encodes the mRuby2 fluorescent protein (Part3) without any signal
            peptide assembled with S. cerevisiae pTEF1 promoter, tTDH1
            terminator, LEU2 selectable marker and CEN/ARS yeast origin.
FEATURES             Location/Qualifiers
     misc_feature    5..10
                     /label="EcoRI Site"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    20..162
                     /label="Con1"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     misc_feature    170..194
                     /label="Con1 scar"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     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="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     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..1617
                     /label="mRuby2"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             905..1717
                     /label="mRuby2"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMRIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGVVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHMALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVAKFAGLGGGMDELYKGSDYKDDDDKDYKDDDDKDYKDDDDKGHHHHHH*"
     misc_feature    1618..1723
                     /label="3XFLAG6XHis"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    1724..1951
                     /label="tTDH1"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     terminator      1728..1951
                     /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    complement(1956..1959)
                     /label="BsmBI"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     protein_bind    complement(1961..1966)
                     /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    1967..2109
                     /label="Con2"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     promoter        2129..2776
                     /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             2777..3871
                     /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      3872..4045
                     /label="AgLEU2 Terminator"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     rep_origin      4050..4166
                     /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      4167..4557
                     /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    4562..4569
                     /label="NotI site"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     terminator      complement(4570..4666)
                     /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(4667..5524)
                     /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(5525..5655)
                     /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(5656..6419)
                     /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    6420..6427
                     /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 atctatggtg tccaaaggag aggagttaat caaggaaaac atgagaatga aagttgtcat
      961 ggagggctcc gttaatggtc accaattcaa gtgtacaggg gaaggtgaag gtaatcctta
     1021 catgggtaca caaactatga gaattaaagt aattgaaggc ggaccactac catttgcatt
     1081 tgacattctg gcaacgtcat tcatgtacgg atcacgaact ttcatcaagt accctaaagg
     1141 tataccagac tttttcaagc aatcttttcc agagggtttt acatgggaaa gggttacaag
     1201 atacgaagat gggggtgtcg tcacagttat gcaagatact tcattagaag atggctgcct
     1261 tgtctatcat gtgcaagtaa gaggggtgaa ttttccttct aacggacctg tgatgcagaa
     1321 aaagaccaaa ggttgggaac caaatactga aatgatgtac ccagctgatg gaggtttgag
     1381 aggctacaca cacatggcgc ttaaagttga tggtggaggt catttgtctt gtagttttgt
     1441 taccacttat cgttctaaaa agactgttgg caatatcaaa atgccaggaa tacatgctgt
     1501 agaccacaga ctagaaagac tcgaagagag cgataacgaa atgttcgttg tacagagaga
     1561 gcatgccgta gccaaatttg ctggcttagg cggtggtatg gatgaattgt ataagggatc
     1621 cgattacaag gacgatgatg ataaggacta caaagatgat gatgacaaag attataagga
     1681 cgatgatgat aaaggtcatc atcatcatca tcactaactc gagtggcata aagcaatctt
     1741 gatgaggata atgatttttt tttgaatata cataaatact accgtttttc tgctagattt
     1801 tgtgatgacg taaataagta catattactt tttaagccaa gacaagatta agcattaact
     1861 ttaccctttt ctttctaagt ttcaatatta gttatcactg tttaaaagtt atggcgagaa
     1921 cgtcggcggt taaaatatat taccctgaac ggctggatgt gagacgataa atcgcagcca
     1981 agtgagtgaa tagatgacgc accacggtca gacacggcca catcgtatct cacaggagca
     2041 agcgcgatag gagcactcac acatagtacg gtgatccgct gactcctttg cccaaataag
     2101 acgtgagcca ctagtgcact gcagtacatc gaggagaact tctagtatat ctacatacct
     2161 aatattattg ccttattaaa aatggaatcc caacaattac atcaaaatcc acattctctt
     2221 caaaatcaat tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta tatttatagg
     2281 ataattatac tctatttctc aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc
     2341 tgattcaaga aatatcttga ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc
     2401 aagagttcga atctcttagc aaccattatt tttttcctca acataacgag aacacacagg
     2461 ggcgctatcg cacagaatca aattcgatga ctggaaattt tttgttaatt tcagaggtcg
     2521 cctgacgcat ataccttttt caactgaaaa attgggagaa aaaggaaagg tgagagcgcc
     2581 ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata
     2641 cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc
     2701 gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat atatatttca
     2761 aggatatacc attctaatgt ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt
     2821 tggtcaagaa atcacagccg aagccattaa ggttcttaaa gctatttctg atgttcgttc
     2881 caatgtcaag ttcgatttcg aaaatcattt aattggtggt gctgctatcg atgctacagg
     2941 tgttccactt ccagatgagg cgctggaagc ctccaagaag gctgatgccg ttttgttagg
     3001 tgctgtgggt ggtcctaaat ggggaaccgg tagtgttaga cctgaacaag gtttactaaa
     3061 aatccgtaaa gaacttcaat tgtacgccaa cttaagacca tgtaactttg catccgactc
     3121 tcttttagac ttatctccaa tcaagccaca atttgctaaa ggtactgact tcgttgttgt
     3181 cagagaatta gtgggaggta tttactttgg taagagaaag gaggacgatg gtgatggtgt
     3241 cgcttgggat agtgaacaat acaccgttcc agaagtgcaa agaatcacaa gaatggccgc
     3301 tttcatggcc ctacaacatg agccaccatt gcctatttgg tccttggata aagctaatgt
     3361 tttggcctct tcaagattat ggagaaaaac tgtggaggaa accatcaaga acgaatttcc
     3421 tacattgaag gttcaacatc aattgattga ttctgccgcc atgatcctag ttaagaaccc
     3481 aacccaccta aatggtatta taatcaccag caacatgttt ggtgatatca tctccgatga
     3541 agcctccgtt atcccaggtt ccttgggttt gttgccatct gcgtccttgg cctctttgcc
     3601 agacaagaac accgcatttg gtttgtacga accatgccac ggttctgctc cagatttgcc
     3661 aaagaataag gtcaacccta tcgccactat cttgtctgct gcaatgatgt tgaaattgtc
     3721 attgaacttg cctgaagaag gtaaggccat tgaagatgca gttaaaaagg ttttggatgc
     3781 aggtatcaga actggtgatt taggtggttc caacagtacc accgaagtcg gtgatgctgt
     3841 cgccgaagaa gttaagaaaa tccttgctta acctggagga cccttctctt tagactattc
     3901 tactcttatg cacgtaaaaa attctaggaa atatgtatta actaggagta aaataaccgg
     3961 ctagtggcat tcatatagcc gtctgtttac atctacatca cacatttcga gtgtatatct
     4021 cgcaacgttg gcgttaaata ggcaggagta tcacgtgcta taaaaataat tataatttaa
     4081 attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa
     4141 atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct
     4201 tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta
     4261 gaaaaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata
     4321 tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa
     4381 attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat
     4441 ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa
     4501 attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atccgtcccg
     4561 agcggccgcg attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4801 cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5641 gaaaagtgcc acctgtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6361 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
     6421 cggccgc
//