LOCUS       pLigify_Isoeugenol      4505 bp ds-DNA     circular     23-JAN-2024
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    complement(61..83)
                     /label="GFP RBS8 450K"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(84..161)
                     /label="ElvJ"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    complement(162..335)
                     /label="iem promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    336..384
                     /label="Bidirectional terminator"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    392..444
                     /label="P250"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     ribozyme        457..531
                     /label="riboJ"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    532..563
                     /label="5'-UTR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    564..614
                     /label="Leader peptide"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             564..614
                     /label="Translation 564-614"
                     /translation="MKAIFVLKHLNHAPVF*"
     misc_feature    602..610
                     /label="4.6K"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    616..1566
                     /label="iemR"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             616..1566
                     /label="imeR"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /translation="MTTIRWRRMSIHSERITLADSPLHWAHTLNGSMRTHFEVQRLERGRGAYLARSRFGAGELYSAIAPSQVLRHFNDQRNANEAEHSYLIQIRSGALGVASGGRKVILANGDCSIVDSRQDFTLSSNSSTQGVVIRFPVSWLGAWVSNPEDLIARRVDAEIGWGRALSASVSNLDPLRIDDLGSNVNSIAEHVAMLISLASSAVSSEDGGVALRKMREVKRVLEQSFADANLEPESVSSQLGISKRYLHYVFAACGTTFGRELLEIRLGKAYRMLCATSGSGAVLKVAMSSGFSDSSHFSKKFKERYGVSPVSLVRQA*"
     misc_feature    1567..3769
                     /label="pTwist Kan Medium Copy"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1575..1664
                     /label="ECK12_spy_term"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     rep_origin      complement(1917..2462)
                     /label="p15A ori"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="direction: LEFT note: Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin."
     misc_feature    complement(2573..3388)
                     /label="KanR"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     terminator      3526..3572
                     /label="rrnB T1 terminator"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="note: transcription terminator T1 from the E. coli rrnB gene"
     misc_feature    complement(3770..3832)
                     /label="L3S2P00"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             complement(3849..60)
                     /label="GFPmut2"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             complement(3849..60)
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK*"
ORIGIN
        1 accatctaat tcaacaagaa ttgggacaac tccagtgaaa agttcttctc ctttactcat
       61 ATATACCCCC TTATTCTCCC GTATTAAACA AAATTATTTG TAGAGGCCCC ATTTCGTCCT
      121 TTTGGACTCA TCAGGGGTGG TACACACCAC CCTATGGGGC TGGCACGGTC TCCTTTGTTG
      181 TTCTGAATGG CCTAAATGCG CGGCTTGCCG GGTTGGAGTT TATGTTTAGG ACTGACCGGA
      241 TTTCATGTGT GCCGGTGAAG TGAGGATGTC TGTGAGTGCA ATGGTGGCGG TATTGAAAAT
      301 GGGCCGAGGC TGGCCTATTG TTGAGGATTT CAAGAtttct tgcaaaaaaa aaccccgccg
      361 aagcggggtt ttttttttta caggggAAAC Gaaaatatat ttttcaaaag tatcgTTTAC
      421 Cgctagctca gtcctaggTA CAATgagcac agTGGCagct gtcaccggat gtgctttccg
      481 gtctgatgag tccgtgagga cgaaacagcc tctaCAAATA ATTTTGTTTA AGGGCCCAAG
      541 TTCACTTAAA AAGGAGATCA ACAATGAAAG CAATTTTCGT ACTGAAACAT CTTAATCATG
      601 CGCCGGTGTT TTAAAATGAC CACCATCCGC TGGCGCCGTA TGTCTATTCA TTCGGAGCGC
      661 ATCACTCTTG CAGATAGCCC TTTACATTGG GCACATACAT TGAATGGTTC TATGCGTACC
      721 CATTTTGAAG TACAACGCTT AGAGCGCGGG CGCGGTGCTT ACTTGGCGCG CAGTCGCTTC
      781 GGTGCTGGGG AATTGTACTC GGCCATCGCC CCTTCCCAGG TCTTACGTCA CTTCAATGAT
      841 CAACGTAACG CCAACGAAGC GGAACACTCC TACCTGATTC AGATCCGCTC GGGGGCCTTA
      901 GGCGTGGCCA GTGGAGGTCG TAAAGTTATC TTAGCAAATG GCGATTGTAG TATTGTCGAT
      961 TCGCGCCAAG ACTTCACGTT GTCGTCCAAC TCATCTACTC AAGGGGTCGT GATCCGTTTT
     1021 CCTGTGAGTT GGTTGGGTGC CTGGGTATCC AATCCCGAAG ATCTTATTGC CCGTCGCGTT
     1081 GACGCAGAAA TTGGATGGGG GCGCGCGTTG TCTGCTAGTG TCTCCAACTT AGACCCGCTG
     1141 CGTATCGACG ACTTAGGAAG TAACGTCAAT TCGATTGCAG AGCATGTCGC TATGTTAATT
     1201 TCACTTGCTA GCTCGGCTGT GAGTTCGGAG GACGGGGGAG TAGCCTTACG CAAAATGCGT
     1261 GAGGTCAAGC GCGTCTTAGA ACAGTCATTT GCCGATGCGA ATCTTGAACC AGAAAGTGTT
     1321 TCAAGCCAGC TTGGTATTAG TAAGCGCTAC TTACACTACG TTTTTGCTGC GTGTGGAACA
     1381 ACTTTTGGCC GTGAATTATT GGAGATCCGC TTGGGAAAGG CTTACCGTAT GCTGTGTGCG
     1441 ACTTCAGGAA GTGGGGCTGT TCTTAAGGTG GCTATGTCCA GTGGGTTTAG CGATTCGAGC
     1501 CACTTCTCCA AAAAATTCAA GGAACGTTAC GGAGTGTCTC CTGTGTCGTT AGTCCGCCAA
     1561 GCTTAATAAT CACTTTCAGC CAAAAAACTT AAGACCGCCG GTCTTGTCCA CTACCTTGCA
     1621 GTAATGCGGT GGACAGGATC GGCGGTTTTC TTTTCTCTTC TCAAaggcta ggtggaggct
     1681 cagtgatgat aagtctgcga tggtggatgc atgtgtcatg gtcatagctg tttcctgtgt
     1741 gaaattgtta tccgctcaga gggcacaatc ctattccgcg ctatccgaca atctccaaga
     1801 cattaggtgg agttcagttc ggcgagcgga aatggcttac gaacggggcg gagatttcct
     1861 ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc
     1921 cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga
     1981 aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct
     2041 cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat
     2101 tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga
     2161 accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2221 ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag
     2281 tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc
     2341 ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc
     2401 cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc
     2461 aagaagatca tcttattaag tctgacgctc tattcaacaa agccgccgtc catgggtagg
     2521 gggcttcaaa tcgtccgctc tgccagtgtt acaaccaatt aacaaattct gattagaaaa
     2581 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     2641 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     2701 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     2761 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     2821 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     2881 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     2941 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     3001 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     3061 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     3121 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     3181 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     3241 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     3301 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc
     3361 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     3421 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     3481 gagacacaac gtggctttcc cccgccgctc tagaactagt ggatccaaat aaaacgaaag
     3541 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcgcatta tacgagacgt
     3601 ccaggttggg atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt
     3661 gatccaccac aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat
     3721 gcataatccg cacgcatctg gaataaggaa gtgccattcc gcctgacctg gaccaaaacg
     3781 aaaaaagggg agcggtttcc cgctcccctc ttttctggaa tttggtaccg agGAATGAAg
     3841 CAGGATtatt atttgtatag ttcatccatg ccatgtgtaa tcccagcagc tgttacaaac
     3901 tcaagaagga ccatgtggtc tctcttttcg ttgggatctt tcgaaagggc agattgtgtg
     3961 gacaggtaat ggttgtctgg taaaaggaca gggccatcgc caattggagt attttgttga
     4021 taatggtctg ctagttgaac gcttccatct tcaatgttgt gtctaatttt gaagttaact
     4081 ttgattccat tcttttgttt gtctgccatg atgtatacat tgtgtgagtt atagttgtat
     4141 tccaatttgt gtccaagaat gtttccatct tctttaaaat caataccttt taactcgatt
     4201 ctattaacaa gggtatcacc ttcaaatttg acttcagcac gtgtcttgta gttcccgtca
     4261 tctttgaaaa atatagttct ttcctgtaca taaccttcgg gcatggcact cttgaaaaag
     4321 tcatgctgtt tcatatgatc tgggtatctc gcaaagcatt gaagaccata cgcgaaagta
     4381 gtgacaagtg ttggccatgg aacaggtagt tttccagtag tgcaaataaa tttaagggta
     4441 agttttccgt atgttgcatc accttcaccc tctccactga cagaaaattt gtgcccatta
     4501 acatc
//