LOCUS       sequence-321966-        7683 bp    DNA     circular SYN 15-DEC-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 7683)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 7683)
  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: pEGB3_alpha1 Pnos:P19:Tnos (GB2762)
FEATURES             Location/Qualifiers
     source          1..7683
                     /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="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3715..4788
                     /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      4854..5048
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5284..5306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5392..5532
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5447..5466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5547..5564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5718..6306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5798..5817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6394..7209)
                     /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(6394..7209)
                     /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     7117..7136
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    7642..7666
                     /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 tcgcgcgcgg 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 gcgatgagta aataaacaaa
     2641 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     2701 gacgaccatc gcaacccatc 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 catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     3541 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     3601 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     3661 ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     3721 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     3781 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     3841 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     3901 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     3961 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     4021 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     4081 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     4141 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     4201 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     4261 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     4321 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     4381 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     4441 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     4501 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     4561 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     4621 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     4681 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     4741 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     4801 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggactctt tcctgtggat
     4861 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     4921 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     4981 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     5041 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
     5101 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     5161 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     5221 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     5281 aaacctctga cacatgcagc tcccggtgac ggtcacagct tgtctgtaag cggatgccgg
     5341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     5401 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     5461 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5521 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5581 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5641 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5701 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5761 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5821 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5881 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5941 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6001 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6061 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6121 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6181 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6241 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6301 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6361 cgttaaggga ttttggtcat gcattctagg tgattagaaa aactcatcga gcatcaaatg
     6421 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     6481 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     6541 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     6601 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     6661 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     6721 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagtcgaa atacgcgatc
     6781 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     6841 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     6901 ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     6961 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     7021 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     7081 caatcggtag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     7141 taaatcagca tccatgttgg aatttaatcg cggccttgag caagacgttt cccgttgaat
     7201 atggctcata acagaactta ttatttcctt cctcttttct acagtattta aagatacccc
     7261 aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc taaaacctta
     7321 aataccagaa aacagctttt tcaaagttgt tttcaaagtt ggcgtataac atagtatcga
     7381 cggagccgat tttgaaaccg cggtgatcac aggcagcaac gctctgtcat cgttacaatc
     7441 aacatgctac cctccgcgag atcatccgtg tttcaaaccc ggcagcttag ttgccgttct
     7501 tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt acaacggctc tcccgctgac
     7561 gccgtcccgg actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
     7621 ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacataa cgaattcgtc
     7681 tca
//
