LOCUS       sequence-321955-        7021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pEGB3a2 AtS/MAR10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7021)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 7021)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    665..689
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1990..2619
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3048..4121
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     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 agcttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc
      721 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac
      781 cacagggttc ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag
      841 tcggcttctg acgttcagtg cagccgtcat ctgaaaacga catgtcgcac aagtcctaag
      901 ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc
      961 gcataaagta gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg
     1021 ccgctggcct gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac
     1081 gggccgaact gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca
     1141 ggcgcgaccg cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga
     1201 cagtgaccag gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc
     1261 gcatccagga ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca
     1321 cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc
     1381 taatcatcga ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg
     1441 gcccccgccc taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg
     1501 aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc
     1561 gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc
     1621 gtgaggacgc attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg
     1681 aacaagcatg aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat
     1741 cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacc tctcaaccgt
     1801 gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag
     1861 cttggccgct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa
     1921 cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa
     1981 ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc
     2041 atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat
     2101 tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc
     2161 gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac
     2221 ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca
     2281 gccgacttcg tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac
     2341 ctggtggagc tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt
     2401 gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc
     2461 gggtacgagc tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact
     2521 gccgccgccg gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc
     2581 caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa
     2641 tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg
     2701 caacgttggc cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg
     2761 cggaggatca caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc
     2821 tgctatctga atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga
     2881 tgaattttag cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc
     2941 gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgcc
     3001 tgccggccct gcaatggcac tggaaccccc aagcccgagg aatcggcgtg agcggtcgca
     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
//
