LOCUS       sequence-311000-        7021 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  pEGB3a2 AtS/MAR10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7021)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 7021)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AtS/MAR10 insulator sequence (GB3459)
FEATURES             Location/Qualifiers
     source          1..7021
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    666..690
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1991..2620
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3054..4121
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      4187..4381
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4617..4639)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4725..4865
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4780..4799
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4880..4897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5051..5639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5131..5150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5727..6542)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5727..6542)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6450..6469
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6975..6999
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagtggcta ttgttgttat catcaaaatc tatattgata ctattatact cgatatgtta
       61 tagactttat taaaaaaaac tttgtttgtt tgcagaaagt ggatttttaa tcaaatttaa
      121 ttgtgatgat tacaatgtca attggttatg caagatatta attaatatgt ttaagatttt
      181 tcacttttaa taagtaaaat aaaatacgtc aaaaaaattt cctaaagata atgccaaaaa
      241 ggttaaatta tagaaattaa actaaaatat attaaaagtt gacttaaaaa tgcaatgttg
      301 accaccgacc taggggaaaa caaatttacc gtatctggga tagaatctct tctgtatcat
      361 ctcagataaa gattactaca taaacacact ttacagtttt acgacgccca aaattcaatt
      421 cccggagatt aaaaacccta aaacaaacac tggcgattcg attttccctc ccaattcaaa
      481 tcatctctta atcttcttct tctcactatc ctctcaaatt cgccgccgat cttccatcaa
      541 ttttgatttc tcgcaccgaa atttcgactt ttcatctcct atcgacgaaa ccctaatttt
      601 tcacactcca gaaatcgaac tttgacgacg atgttaatgg ctaaaccccg cttgagacga
      661 agctttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat
      721 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa
      781 ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca
      841 gtcggcttct gacgttcagt gcagccgtca tctgaaaacg acatgtcgca caagtcctaa
      901 gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt
      961 cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc
     1021 gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa
     1081 cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc
     1141 aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg
     1201 acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag
     1261 cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc
     1321 acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc
     1381 ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt
     1441 ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag
     1501 gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac
     1561 cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc
     1621 cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag
     1681 gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga
     1741 tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac ctctcaaccg
     1801 tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca
     1861 gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa
     1921 acagcttgcg tcatgcggtc gctgcgtata tgatccgatg agtaaataaa caaatacgca
     1981 aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac
     2041 catcggaacc catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga
     2101 ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac
     2161 cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga
     2221 cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc
     2281 agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga
     2341 cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt
     2401 tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc
     2461 cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac
     2521 tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt
     2581 ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa
     2641 atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct
     2701 gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg
     2761 gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag
     2821 ctgctatctg aatagatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag
     2881 atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc
     2941 cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgt
     3001 ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt gacggtcgca
     3061 aaccatccgg cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga
     3121 aggccgcgca ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt
     3181 ggcaagcggc cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc
     3241 cgtcgattag gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct
     3301 atgacgtggg cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga
     3361 agcgtgaccg acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg
     3421 tttccgcagg gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg
     3481 tttcccatct aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc
     3541 gcgtgttccg tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa
     3601 agcagaaaga cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc
     3661 agcgtacgaa gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga
     3721 ttagccgcta caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc
     3781 tagctgattg gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc
     3841 accccgatta ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc
     3901 gcgccgcagg caaggcagaa gccagatggt tgttcaagac gatctacgaa cgcagtggca
     3961 gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc
     4021 tgccggagta cgatttgaag gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct
     4081 accgcaacct gatcgagggc gaagcatccg ccggttccta atgtacggag cagatgctag
     4141 ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggact ctttcctgtg gatagcacgt
     4201 acattgggaa cccaaagccg tacattggga accggaaccc gtacattggg aacccaaagc
     4261 cgtacattgg gaaccggtca cacatgtaag tgactgatat aaaagagaaa aaaggcgatt
     4321 tttccgccta aaactcttta aaacttatta aaactcttaa aacccgcctg gcctgtgcat
     4381 aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc
     4441 gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg
     4501 ctggcctacg gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc
     4561 gccggcgccc acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4621 tgacacatgc agctcccggt gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4681 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     4741 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     4801 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4861 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5461 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5701 ggattttggt catgcattct aggtgattag aaaaactcat cgagcatcaa atgaaactgc
     5761 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     5821 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     5881 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     5941 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     6001 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     6061 accaaaccgt tattcattcg tgattgcgcc tgagcgagtc gaaatacgcg atcgctgtta
     6121 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     6181 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttccctggg
     6241 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     6301 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     6361 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcgg
     6421 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     6481 gcatccatgt tggaatttaa tcgcggcctt gagcaagacg tttcccgttg aatatggctc
     6541 ataacagaac ttattatttc cttcctcttt tctacagtat ttaaagatac cccaagaagc
     6601 taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc ttaaatacca
     6661 gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc
     6721 gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca atcaacatgc
     6781 taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat
     6841 agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc
     6901 cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
     6961 tgttggctgg ctggtggcag gatatattgt ggtgtaaaca taacaagctt cgtctcagtc
     7021 a
//
