LOCUS       pCENTR_DMC1p_ami        8095 bp DNA     circular SYN 25-JUN-2024
FEATURES             Location/Qualifiers
     source          1..8095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2946..6081
                     /chromosome="3"
                     /map="between 560B2 and g4711"
                     /ecotype="Landsberg erecta"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:3702"
                     /organism="Arabidopsis thaliana"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2064..2141
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2142..2492
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSG
                     DGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAVANCRGD*ISRRSTGCQRGGVDGR
                     TKRRRSL*SGGAQ"
     terminator      2530..2557
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2649..2735
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2799..2815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2844..2943
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2946..6081
                     /label=AtDMC1_Promoter
                     /note="Klimjuk 1997 sequence of LER DMC1
                     and Lucia Barra 2021 primers"
     gene            5999..6081
                     /gene="AtDMC1"
                     /label=AtDMC1
     mRNA            5999..6081
                     /gene="AtDMC1"
                     /product="AtDMC1"
                     /label=AtDMC1 mRNA
                     /note="recA-like protein"
     CDS             6079..6081
                     /codon_start=1
                     /gene="AtDMC1"
                     /product="AtDMC1"
                     /label=AtDMC1
                     /note="recA-like protein"
                     /protein_id="AAC49617.1"
                     /translation="M"
     misc_feature    6082..7992
                     /label=Region needed for miRNA cloning
     misc_feature    6082..6106
                     
     promoter        join(6355..6360,6361..6378,6379..6385)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6439..7098
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             7440..7745
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(7994..8093)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    8064..8082
                     /label=F1
ORIGIN
        1 atccagctga taagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
       61 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
      121 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      181 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      241 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      301 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      361 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      421 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
      481 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      541 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      601 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      661 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      721 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
      781 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      841 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      901 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      961 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1021 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1081 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1381 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1921 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1981 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     2041 acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg gtgcaaaacc
     2101 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     2161 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     2221 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     2281 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     2341 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     2401 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     2461 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt accgctagcc
     2521 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     2581 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     2641 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     2701 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     2761 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga
     2821 cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt
     2881 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagcag
     2941 gcttatcagc tatgagatta ctcgtgtatc aattctctaa ttaattaaaa atagtataaa
     3001 ttaaataata tagttcgata cacgaatata attgcgaaga ataggcatac aaatttgtca
     3061 tacatgtttc gatatggctc acgaggaggc tgatgcaaca gtttgatgta tacgtatgca
     3121 aattgagaag tacttgatca gacctatata tgtgatgctc gaacttatct ttttgttttg
     3181 gatcatctat cgaatacaat ggtactataa tttaaatgtt ttttttcttc tttttcttta
     3241 gtatcaaaag caacgttaga tgctaaataa agagttagtt gattgtgatg actgatagtc
     3301 tgataatatc attaactttg cacccgaagt caaataaaag tgttcatatt tataaattcc
     3361 aaccaacgtt aataagccac acctaatcgg tgattgccaa caatattata ataaaattaa
     3421 aaaaactacg actaaagtta atttgctata attttgtggt atgttttaaa aataaagttc
     3481 tttagttcta atatcatgaa aattcagtgt actgtaaaat atgtaaaaag gttttagtac
     3541 aattcttttt tgtatataac ggcaaagttc aatacatatt ttactattga ttttttttaa
     3601 aaataaaata acaattgcta ccaacttttt gaagcatatt gatcgcaact taattataat
     3661 tcttcttttt tttcttggaa gattaataaa acctaatttc aatgtggaac aaataaatgt
     3721 agaaatattg ttatcacaaa ctaatatatg atatttttta atattttcat atatactttt
     3781 gagcttctga tgatataaca gttttcatta aaatacaaat tgtcgtgtac taatttttct
     3841 tttgttcaag tatgtgataa aaatatgttg caaaattgcg agttattata atggtacaaa
     3901 tatgtagaga gaatacatga gaagagttaa aagaagcatg cttaagccaa cagagagtgg
     3961 atccaaatgt tgctttccag ctttatacaa acgtatcacc cacattactg ccactgctac
     4021 atatattgaa ggagagagag atgatgatca taatgataaa tcgatgtcga tgataaattg
     4081 atgatgatgg ctccggtatg tgtaccctag gagttgtagc tagctagcta ggaccatgta
     4141 tatacataca tacatatatt agtgtttttt gtaacttgta cgtaccttac aaacagtatg
     4201 gagtttacta aaacggcaac gtttggtggg ggtagtgaat tcgcaagtgg ggatgagtct
     4261 atgtaataga agatgcaaca tgcaaatggt cccctttctg tttttattta aagaaattag
     4321 tgtttactga ggaggaaaca tcccatttat agattcacac ccataaaagc aaaccacttc
     4381 tccttctttt tatttcccca tgatattact tcgagaatat tttgaaaatt tgaagtgtac
     4441 atttagagat tgtgtacttt gaacactcat gtcaaatgca tctaaatata taaactccaa
     4501 tttaaaataa tcgtctaaac ctagagtgcc atttgtttag ccatttgttg gtcttcattt
     4561 ctcatgcttt gattacatgt accggttgat tcatgtgaaa aatcatgtgc ataaactaag
     4621 aaatagctag cacataaaat tttgatttag gttggatatt actatgttca ctttaagaga
     4681 aaaaaaaact tatggcaaaa agtgatgatg gtatatgaat atgataatca aagtgcatat
     4741 gtgaagtgag aggcaactgt agagtaatat aataaaatcc aaagaaaatt tttaaatatg
     4801 agaaaaaatt atataaaaag gttcttttgt aatccacttc ttttgatata gggagattcg
     4861 ttgagcatcc atgtgctctt tcaatcgaca ctattctgtc tgtatctagc caacccacat
     4921 atacctttac actagagaac ttcgatgatt ctttttccaa aatcaatgtg atataatata
     4981 attaagcata tatgcataaa aaatgaagaa gaatggtaga gtcatgttac ttaaggtcat
     5041 ggtgtgtaaa aacattgata ctttacaata tatgagttgt gaagtgctct taaagttata
     5101 acatccggtt ctacgtattg acctagaact agaagaatcg ttttttagtc caaatcaaat
     5161 caagtcggtt ctttatcagt tttgttgtat gtgaattaat ttgaaaatat tagctatgat
     5221 cttagcttgg gtttttgttt ctaagggtta aggatcatat ctctttgtca aatgacatgt
     5281 ggtctatatg tcatgaatta ggcaccgcta tcttttacta ttgattcgac gacattggga
     5341 ctcctcacta cacttatctt aaaaaaactc aaagttggtg ttaatggctt gtcaccataa
     5401 actttcatga gctctaacaa attaaacttg aacttgatca ggtctcacaa tatatacaat
     5461 ttcgagggat aaatatttca aaaggataat atgatagttg gtagaaatgt atagtttcta
     5521 gtaataatag agatcgttgg ttaaactccc caacttttta aaattaattt gattagtgga
     5581 tccgcaaaca aatattagat tgggcctata tgcatctata ttatttttat ttttctgtaa
     5641 tttcagtaaa atgggcctat ggtcctatat gcatccgaat aattagtata ctgggcttat
     5701 gggcctatat gcatttgatt ttatcgataa aatgtgagtc aaatgtctaa tgtgcgccgt
     5761 tatgaagtgc aagtggctaa tttttttcac ctagattcct tctattgacc gtcgatagac
     5821 ggatgataac tatgacgtgg cattatcgca gccatcaaac aaagtcatgt ataacaaaca
     5881 agagcacaca aacgaaaaca aattcagttg cggaacccaa attcaaatca acggaattag
     5941 aatcacgctt tcaattccgt aacccgccat taaaaacctt gaaccctcga agcaaatcga
     6001 gcaaagattt tcaaatttcg aatttcaaaa ttctatctct ctcactcttc caagcttaga
     6061 gagtcttaga gcgagaaaat gtataggggg gaaaaaaagg tagtcatcag atatatattt
     6121 tggtaagaaa atatagaaat gaataatttc acgtttaacg aagaggagat gacgtgtgtt
     6181 ccttcgaacc cgagttttgt tcgtctataa atagcacctt ctcttctcct tcttcctcac
     6241 ttccatcttt ttagcttcac tatctctcta taatcggttt tatctttctc taagtcacaa
     6301 cccaaaaaaa caaagtagag aagaatctgt aagagaccat taggcacccc aggctttaca
     6361 ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggagccgtc gagattttca
     6421 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     6481 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     6541 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     6601 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggagttccgt
     6661 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     6721 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     6781 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     6841 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     6901 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     6961 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     7021 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     7081 tggcagggcg gggcgtaaac gcgtggagcc ggcttactaa aagccagata acagtatgcg
     7141 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     7201 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     7261 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     7321 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     7381 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
     7441 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     7501 agagtgatat tattgacacg cccggccgac ggatggtgat ccccctggcc agtgcacgtc
     7561 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     7621 ggcgcatgat gaccaccgat atggccagtg tgccggtttc cgttatcggg gaagaagtgg
     7681 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     7741 tataaatgtc aggctccctt atacacagcc agtctgcacc tcgacggtct cacattggct
     7801 cttcttacta caatgaaaaa ggccgaggca aaacgcctaa aatcacttga gaatcaattc
     7861 tttttactgt ccatttaagc tatcttttat aaacgtgtct tattttctat ctcttttgtt
     7921 taaactaaga aactatagta ttttgtctaa aacaaaacat gaaagaacag attagatctc
     7981 atctttagtc tcaacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     8041 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgcc
//
