LOCUS       Exported                9960 bp ds-DNA     circular SYN 27-OCT-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9960)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2022 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9960
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..1343
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
                     /note="color: #ffff00"
     promoter        1349..1564
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1565..2368
                     /label=URA3
                     /note="color: #993366; direction: RIGHT"
     misc_feature    2339..5667
                     /label=YRC Fragment: Q5_At_DNG02774_2
                     /label=Fragment 2
                     /note="color: #ff0000"
     rep_origin      2368..2998
                     /label=oriV
                     /note="incP origin of replication"
                     /note="color: #ffff00"
     CDS             3210..4004
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /note="color: #ccffcc"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             4303..5451
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /note="color: #993366"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    5603..5627
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
                     /note="color: #ffe4c4"
     misc_feature    5637..7195
                     /label=YRC Fragment: Q5_At_DNG02774_3
                     /label=Fragment 4
                     /note="color: #ff0000"
     misc_feature    5668..9803
                     /label=Replacement Cassette
                     /note="color: #0000ff"
     misc_feature    5668..7167
                     /label=Homology Region DNG_02774
                     /label=HU Region of DNG_02774
                     /note="color: #33cccc"
     misc_feature    7168..8603
                     /label=YRC Fragment: Q5_At_DNG02774_4
                     /note="color: #ff0000"
     promoter        7168..7549
                     /label=trpC-P
                     /note="color: #ffffff"
     CDS             7544..8569
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /note="color: #ccffcc"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    8570..8603
                     /label=T.Trpc
                     /note="color: #a6acb3"
     misc_feature    8581..9833
                     /label=YRC Fragment: Q5_At_DNG02774_5
                     /label=Fragment 5
                     /note="color: #ff0000"
     misc_feature    8604..9803
                     /label=Homology Region DNG_02774
                     /label=HD region DNG_02774
                     /note="color: #33cccc"
     misc_feature    join(9804..9960,1..2394)
                     /label=YRC Fragment: Q5_At_DNG02774_1
                     /label=Fragment1
                     /note="color: #ff0000"
     repeat_region   9804..9960
                     /label=T-DNA LB
                     /label=nonstandard type: Repeat_unit
                     /note="color: #a6acb3"
     misc_feature    9857..9881
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
                     /note="color: #ffe4c4"
ORIGIN
        1 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
       61 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
      121 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
      181 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
      241 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
      301 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
      361 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
      421 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
      481 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
      541 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
      601 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
      661 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
      721 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
      781 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
      841 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
      901 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
      961 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
     1021 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
     1081 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
     1141 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
     1201 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta
     1261 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
     1321 atcatttcct ttgatattgg atcttcaatt caattcatca tttttttttt attctttttt
     1381 ttgatttcgg tttctttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg
     1441 aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg
     1501 aaattgccca gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata
     1561 aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc
     1621 caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg
     1681 taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa
     1741 aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc
     1801 attatccgcc aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa
     1861 tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac
     1921 gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga
     1981 agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct
     2041 atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt
     2101 tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat
     2161 tatgacaccc ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac
     2221 cgtggatgat gtggtctcta caggatctga cattattatt gttggaagag gactatttgc
     2281 aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata
     2341 tttgagaaga tgcggccagc aaaactaacg ctcaccgggc tggttgccct cgccgctggg
     2401 ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga
     2461 caccgcggcc gccggcgttg tggatacctc gcggaaaact tggccctcac tgacagatga
     2521 ggggcggacg ttgacacttg aggggccgac tcacccggcg cggcgttgac agatgagggg
     2581 caggctcgat ttcggccggc gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc
     2641 tgattttacg cgagtttccc acagatgatg tggacaagcc tggggataag tgccctgcgg
     2701 tattgacact tgaggggcgc gactactgac agatgagggg cgcgatcctt gacacttgag
     2761 gggcagagtg ctgacagatg aggggcgcac ctattgacat ttgaggggct gtccacaggc
     2821 agaaaatcca gcatttgcaa gggtttccgc ccgtttttcg gccaccgcta acctgtcttt
     2881 taacctgctt ttaaaccaat atttataaac cttgttttta accagggctg cgccctgtgc
     2941 gcgtgaccgc gcacgccgaa ggggggtgcc cccccttctc gaaccctccc ggcccgctct
     3001 cgagttggca gcatcaccca taattgtggt ttcaaaatcg gctccgtcga tactatgtta
     3061 tacgccaact ttgaaaacaa ctttgaaaaa gctgttttct ggtatttaag gttttagaat
     3121 gcaaggaaca gtgaattgga gttcgtcttg ttataattag cttcttgggg tatctttaaa
     3181 tactgtagaa aagaggaagg aaataataaa tggctaaaat gagaatatca ccggaattga
     3241 aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct cctgctaagg
     3301 tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac agccggtata
     3361 aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg gaaggaaagc
     3421 tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg ctggagcaat ctgctcatga
     3481 gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tgaacaaagc cctgaaaaga
     3541 ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata tcggattgtc
     3601 cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg aataacgatc
     3661 tggccgatgt ggattgcgaa aactgggaag aagacactcc atttaaagat ccgcgcgagc
     3721 tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc cacggcgacc
     3781 tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt gatcttggga
     3841 gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg atcagggagg
     3901 atatcgggga agaacagtat gtcgagctat tttttgactt actggggatc aagcctgatt
     3961 gggagaaaat aaaatattat attttactgg atgaattgtt ttagtaccta gatgtggcgc
     4021 aacgatgccg gcgacaagca ggagcgcacc gacttcttcc gcatcaagtg ttttggctct
     4081 caggccgagg cccacggcaa gtatttgggc aaggggtcgc tggtattcgt gcagggcaag
     4141 attcggaata ccaagtacga gaaggacggc cagacggtct acgggaccga cttcattgcc
     4201 gataaggtgg attatctgga caccaaggca ccaggcgggt caaatcagga ataagggcac
     4261 attgccccgg cgtgagtcgg ggcaatcccg caaggagggt gaatgaatcg gacgtttgac
     4321 cggaaggcat acaggcaaga actgatcgac gcggggtttt ccgccgagga tgccgaaacc
     4381 atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct tccagtccgt cggctcgatg
     4441 gtccagcaag ctacggccaa gatcgagcgc gacagcgtgc aactggctcc ccctgccctg
     4501 cccgcgccat cggccgccgt ggagcgttcg cgtcgtctcg aacaggaggc ggcaggtttg
     4561 gcgaagtcga tgaccatcga cacgcgagga actatgacga ccaagaagcg aaaaaccgcc
     4621 ggcgaggacc tggcaaaaca ggtcagcgag gccaagcagg ccgcgttgct gaaacacacg
     4681 aagcagcaga tcaaggaaat gcagctttcc ttgttcgata ttgcgccgtg gccggacacg
     4741 atgcgagcga tgccaaacga cacggcccgc tctgccctgt tcaccacgcg caacaagaaa
     4801 atcccgcgcg aggcgctgca aaacaaggtc attttccacg tcaacaagga cgtgaagatc
     4861 acctacaccg gcgtcgagct gcgggccgac gatgacgaac tggtgtggca gcaggtgttg
     4921 gagtacgcga agcgcacccc tatcggcgag ccgatcacct tcacgttcta cgagctttgc
     4981 caggacctgg gctggtcgat caatggccgg tattacacga aggccgagga atgcctgtcg
     5041 cgcctacagg cgacggcgat gggcttcacg tccgaccgcg ttgggcacct ggaatcggtg
     5101 tcgctgctgc accgcttccg cgtcctggac cgtggcaaga aaacgtcccg ttgccaggtc
     5161 ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc actacacgaa attcatatgg
     5221 gagaagtacc gcaagctgtc gccgacggcc cgacggatgt tcgactattt cagctcgcac
     5281 cgggagccgt acccgctcaa gctggaaacc ttccgcctca tgtgcggatc ggattccacc
     5341 cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg aagagttgcg aggcagcggc
     5401 ctggtggaac acgcctgggt caatgatgac ctggtgcatt gcaaacgcta gggccttgtg
     5461 gggtcagttc cggctggggg ttcagccagc gctttactga gatctcctgt ggttggcatg
     5521 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct
     5581 aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac actgatagtt
     5641 taaactgaag gcgggaaacg acaatctgac tctcgaagcg atgtcagctc tgctgcatag
     5701 aacgcctgct cctccccgtc gttcgtgagc aaactctcgt cctccttcgt tgctgacttc
     5761 acggtcgata tgagacctcg acagttctca acccacatgc cgttgctgct cagagactgg
     5821 acgcaccact cgtcccaaat cttagagagc gccttgacac tatcgttcag gggccgaaga
     5881 gagatttgaa tctcgatatc tgccgagccc tctgggatga ttagggcttg gctgatgacg
     5941 atgttccgga gcttgaagcc aagaacgctg gccccattgg cagccgcgcg acggttcgcc
     6001 gcttctatcg ccatgcagat atagccagct cccggataca caatattcga ctgaacaaca
     6061 tgatccctga cccaagggat ttcgtcgtga cggatcatgt tcctccatct tggttccagc
     6121 tcagtcgaat ccaccgaggg cgtccccagg aggtcagccc ggggctcaga cttcaggcga
     6181 agttgcttgc tcgtccttga ttctgcccag tagcgcttgg agtggtccca agcaaaaggt
     6241 ggaaggtcga caagcaggtc tggggttatg gtggagttgg ggcggtttat ggcggcgaag
     6301 ctgacttcgt aacctgcctc gaagagagag caagctaact gcatcgtcgt cacaactgcg
     6361 tcttttccac ggtagagagc agaagtgtac gcaatcccac taccactgag cgtcgggtcg
     6421 agcttaatga tctccttgat gggcccggat agggcagcgt gaggcccaat ctcgacgaga
     6481 gtgtcaacat tgagccctgc cgacgagctc ctcggctggc gtttgccgcg ggaatccagg
     6541 caaagagcct tcaaagctga tgcaaactgc acagtcgaga ccatgttgtc gacccagtac
     6601 tggctgccaa gctcgcttcc gtccagcttc ttgcccctga ccgaggaata gaagttgcac
     6661 ttgtctgcgg tagttgtgtt gacttccagg ctggagagcg ctgccagata tgcctccgcg
     6721 acgacctgca tatggtggga gtgataggct acctcaactc gcagcttccg gttgaacacg
     6781 ccatctttct gaagagccac ctgcaattca tccacggcct gcgagtctcc agcaatggta
     6841 acactctgtg ggctgttgac acaagcaacg gtgagcttcc catgggtgac cagtgcaaag
     6901 cgctcctctg cttcttgggg agttaagctg gcagcgatca tggctccctt cagctcaggg
     6961 tgctccgact tgatcgacgc agcgaggatt ccgcggtggt aggatacgag catggcgccc
     7021 tcgtgggtga tggctccaca gcaatatgcg cccgcgatct caccgctcga gtggccaacc
     7081 acggcggacg gccggacacc gcatgtctct agcaggtcga cgagtgccac ctgaatagca
     7141 gtgcagacag gctgggctag agacgcgcga taccgtcgac gttaactgat attgaaggag
     7201 catttttggg cttggctgga gctagtggag gtcaacaatg aatgcctatt ttggtttagt
     7261 cgtccaggcg gtgagcacaa aatttgtgtc gtttgacaag atggttcatt taggcaactg
     7321 gtcagatcag ccccacttgt agcagtagcg gcggcgctcg aagtgtgact cttattagca
     7381 gacaggaacg aggacattat tatcatctgc tgcttggtgc acgataactt ggtgcgtttg
     7441 tcaagcaagg taagtggacg acccggtcat accttcttaa gttcgccctt cctcccttta
     7501 tttcagattc aatctgactt acctattcta cccaagcatc caaatgaaaa agcctgaact
     7561 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     7621 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     7681 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     7741 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     7801 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     7861 cgaactgccc gctgttctcc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     7921 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     7981 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     8041 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     8101 ggactgcccc gaagtccggc acctcgtgca tgcggatttc ggctccaaca atgtcctgac
     8161 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     8221 atacgaggtc gccaacatcc tcttctggag gccgtggttg gcttgtatgg agcagcagac
     8281 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcgcctcc gggcgtatat
     8341 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     8401 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     8461 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc
     8521 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataga gtagatgccg
     8581 accgggaacc agttaacgtt aacattcttt cgggcctgta tgttcgtatg attttacaat
     8641 gatggccaac tcaggctagg atgattttac acgggtgata gctaactgca actcaaggac
     8701 agcgcagcac acaagcttga gtcttataac tggcagagaa caggttacag tcttcctcac
     8761 ggacgtgtcc gaatccactg acaggataac gcgagaatcg tcgttatcaa taggtacttg
     8821 ctagttacct tggagttgat gtaccgcatc ctatcgtggt cgtctcatgc ggagcactta
     8881 taccgataca ccgctttcaa actccagaga cctagatccc ttccatccag gccgtcaccc
     8941 tgcaggagtc acagcgcaga cgacacacac cgccttccaa atccagctgt agcttgatcg
     9001 gagttaggcc aagatattac attcggtctt aggtaagatc cgccagcttt atttgggtaa
     9061 tagtccgaaa tctgactgta ttttgcttgc taagacaccc gcagcatata gggctattat
     9121 tagccaacga tatccttacc cccacctaag ttagtcagga gcggggcata gacagggaca
     9181 agtaatgcag attcgaaata caccgtgttc ttttgccgct catcatattc cgtatgatag
     9241 atacgcgaac ggccgcgtca gcaggttgcc cctgaaaaga aaagaaagag ggtgagacgg
     9301 ggtgtcgcgg aaggatcccg gggtccacaa cggtcgcgca gtgtcacgat gcggtgtcac
     9361 gatgctcttg actcgcatcg actactacca aggcttctcg gaaggacacg gcatcggtgc
     9421 cgtactccga gatgtatgta caccccagcg gtgtgcatac atacgccctg ttcccccgcc
     9481 cacgttgatc gacatctgac accgccccca tgtataggca agtaccacat ccaaccttgt
     9541 ttgtgccgac gcagccgccg ttctcacggg tggctagctc aactgcaggc gcactgtgat
     9601 attgcccctc tccggaagga ttgagggggt attccgagga tcgattgtca atcgggaagt
     9661 ctgatattag gcccaacctc gccaagtgct tcaccactta ccgatagcta attttaccac
     9721 gccatacgag accaatttgt taccacctac caatacctgg cgtgggtgtg catgagacag
     9781 agagccgtct atctcccata attcagtact ttaaaaacgt ccgcaatgtg ttattaagtt
     9841 gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc
     9901 ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca gtccactaga
//
