LOCUS       sequence-311836-        7682 bp    DNA     circular SYN 15-APR-2025
DEFINITION  TU for expression of P19 driven by Nos promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 7682)
  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: pEGB3_alpha1 Pnos:P19:Tnos (GB2762)
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        83..262
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      839..1091
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1126..1144)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1332..1356
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2657..3286
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3720..4787
                     /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      4853..5047
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5283..5305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5391..5531
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5446..5465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5546..5563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5717..6305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5797..5816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6393..7208)
                     /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(6393..7208)
                     /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     7116..7135
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    7641..7665
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagatacat gagaattaag ggagtcacgg tatgaccccc gccgatgacg cgggacaagc
       61 cgttttacgt ttggaactga cagaaccgca acgttgaagg agccactgag ccgcgggttt
      121 ctggagttta atgagctaag cacatacgtc agaaaccatt attgcgcgtt caaaagtcgc
      181 ctaaggtcac tatcagctag caaatatttc ttgtcaaaaa tgctccactg acgttccata
      241 aattcccctc ggtatccaat tagagtctca tattcactct caactcgatc gaggcaatgg
      301 aacgagctat acaaggaaac gacgctaggg aacaagctaa cagtgaacgt tgggatggag
      361 gatcaggagg taccacttct cccttcaaac ttcctgacga aagtccgagt tggactgagt
      421 ggcggctaca taacgatgag actaattcga atcaagataa tccccttggt ttcaaggaaa
      481 gctggggttt cgggaaagtt gtatttaaga gatatctcag atacgacagg acggaagctt
      541 cactgcacag agtccttgga tcttggacgg gagattcggt taactatgca gcatctcgat
      601 ttttcggttt cgaccagatc ggatgtacct atagtattcg gtttcgagga gttagtatca
      661 ccgtttctgg aggctctcga actcttcagc atctctgtga gatggcaatt cggtctaagc
      721 aagaactgct acagcttgcc ccaatcgaag tggaaagtaa tgtatcaaga ggatgccctg
      781 aaggtactga aaccttcgaa aaagaaagcg agtgagcttg gaatggatct tcgatcccga
      841 tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgac
      901 gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat
      961 gacgttattt atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc
     1021 gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcdcgg tgtcatctat
     1081 gttactagat cgggaattgc caagctaatt cttgaagacg aaagggcctc gtgatacgcc
     1141 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1201 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1261 cgctcatgag acaataaccc tgataaatgc ttcaataatg ggaccgactc gcgctgtcat
     1321 gagacgaatt ctgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag
     1381 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca
     1441 tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat
     1501 agtgcagtcg gcttctgacg ttcagtgcag ccgtcatctg aaaacgacat gtcgcacaag
     1561 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
     1621 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
     1681 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
     1741 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
     1801 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
     1861 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
     1921 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
     1981 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
     2041 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
     2101 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
     2161 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
     2221 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
     2281 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
     2341 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
     2401 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacctct
     2461 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
     2521 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     2581 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat ccgatgagta aataaacaaa
     2641 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     2701 gacgaccatc ggaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     2761 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     2821 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     2881 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     2941 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     3001 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     3061 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     3121 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     3181 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     3241 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     3301 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     3361 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     3421 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     3481 accgagctgc tatctgaata gatcgcgcag ctaccagagt aaatgagcaa atgaataaat
     3541 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     3601 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     3661 ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgacg
     3721 gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga
     3781 agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg
     3841 aatcgtggca agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg
     3901 gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga
     3961 tgctctatga cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc
     4021 tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg
     4081 tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga
     4141 tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc
     4201 ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg
     4261 gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg
     4321 ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag
     4381 ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga
     4441 tcgagctagc tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga
     4501 cggttcaccc cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg
     4561 cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca
     4621 gtggcagcgc cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa
     4681 atgacctgcc ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca
     4741 tgcgctaccg caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga
     4801 tgctagggca aattgcccta gcaggggaaa aaggtcgaaa aggactcttt cctgtggata
     4861 gcacgtacat tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc
     4921 caaagccgta cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag
     4981 gcgatttttc cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct
     5041 gtgcataact gtctggccag cgcacagccg aagagctgca aaaagcgcct acccttcggt
     5101 cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa
     5161 aaatggctgg cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac
     5221 tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa
     5281 aacctctgac acatgcagct cccggtgacg gtcacagctt gtctgtaagc ggatgccggg
     5341 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     5401 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     5461 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     5521 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5581 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5641 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5701 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5761 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5821 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5881 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5941 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6001 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6061 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6121 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6241 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6361 gttaagggat tttggtcatg cattctaggt gattagaaaa actcatcgag catcaaatga
     6421 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     6481 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     6541 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     6601 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     6661 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     6721 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagtcgaaa tacgcgatcg
     6781 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     6841 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     6901 cctgggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     6961 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     7021 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     7081 aatcggtaga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     7141 aaatcagcat ccatgttgga atttaatcgc ggccttgagc aagacgtttc ccgttgaata
     7201 tggctcataa cagaacttat tatttccttc ctcttttcta cagtatttaa agatacccca
     7261 agaagctaat tataacaaga cgaactccaa ttcactgttc cttgcattct aaaaccttaa
     7321 ataccagaaa acagcttttt caaagttgtt ttcaaagttg gcgtataaca tagtatcgac
     7381 ggagccgatt ttgaaaccgc ggtgatcaca ggcagcaacg ctctgtcatc gttacaatca
     7441 acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt
     7501 ccgaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct cccgctgacg
     7561 ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg
     7621 gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacataac gaattcgtct
     7681 ca
//
