LOCUS       sequence-156936-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  binary DEST vector for plant expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  TITLE     pAM-PAT-ProUBQ10 GW
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7420)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAM-PAT-ProATE2 GW
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    348..372
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      484..1193
                     /label=oriV
                     /note="incP origin of replication"
     promoter        1254..1358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1359..1427,1428..2219)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1577..1596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2390..2978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2879..2898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3290..3314
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(3408..3660)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(3680..4231)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(4273..4452)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     protein_bind    5693..5817
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5842..5847,5848..5865,5866..5872)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5926..6585
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5992..6011)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6927..7232
                     /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"
     primer_bind     7111..7130
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(7273..7397)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 agcttgttga gaattcatcg cccgggactg ggttaacttc cgatcgattc tagactagtt
       61 ctagagtccg caaaaatcac cagtctctct ctacaaatct atctctctct atttttctcc
      121 agaataatgt gtgagtagtt cccagataag ggaattaggg ttcttatagg gtttcgctca
      181 tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa
      241 taaaatttct aattcctaaa accaaaatcc agtgaccggg cggccgccac cgcggtggag
      301 ggggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat
      361 tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg
      421 tgaaaaggtt tatccgttcg tccatttgta tgtgtacatc accgacgagc aaggcaagac
      481 cgagcgcctt tccgacgctc accgggctgg ttgccctcgc cgctgggctg gcggccgtct
      541 atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cggccgccgg
      601 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
      661 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
      721 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
      781 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
      841 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
      901 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
      961 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
     1021 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
     1081 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
     1141 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
     1201 agtccttgcc attgccggga tcggggcagt aacgggatgg gcgatcagcc cgacaagcta
     1261 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1321 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1381 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1441 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1501 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1561 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1621 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1681 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1741 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1801 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1861 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1921 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1981 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2041 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2101 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2161 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2221 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2281 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2341 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2401 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2461 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2521 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2581 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2641 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2701 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2761 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2821 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2881 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2941 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3001 tacggttcct ggccttttgc tggccttttg ctcacatgga ctctagctag aggatcacag
     3061 gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc tccgcgagat catccgtgtt
     3121 tcaaacccgg cagcttagtt gccgttcttc cgaatagcat cggtaacatg agcaaagtct
     3181 gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg cctgtatcga
     3241 gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat
     3301 attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa
     3361 tgatcgaata ctaacgtctc taccagatat cagcttgcat gccggtcgat ctagtaacat
     3421 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
     3481 gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat
     3541 gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat
     3601 cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc
     3661 tgcttgactc taggggtcat cagatttcgg tgacgggcag gaccggacgg ggcggcaccg
     3721 gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg
     3781 ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt
     3841 cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca
     3901 gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg gagacgtaca
     3961 cggtcgactc ggccgtccag tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga
     4021 tgccggcgac ctcgccgtcc acctcggcga cgagccaggg atagcgctcc cgcagacgga
     4081 cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt acggaagttg accgtgcttg
     4141 tctcgatgta gtggttgacg atggtgcaga ccgccggcat gtccgcctcg gtggcacggc
     4201 ggatgtcggc cgggcgtcgt tctgggctca tggtagatcc cctcgatcga gttgagagtg
     4261 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg
     4321 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
     4381 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca
     4441 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
     4501 cataacgtga ctcccttaat tctccgctca tgatcagatt gtcgtttccc gccttcggtt
     4561 tgggcgcgcc aagcacctaa acattccaaa caacacattc atgtttcaac aactttacat
     4621 tttgtattca aacaaacaaa caatcgagca ttacaatgct ttattaacca aacctggaag
     4681 aacgatgttt gagatgggtc aagaccagtg ttgccaggtt gaacaaccac atcaatagga
     4741 gcaactaaac ccacacgagc aggagcacca acctgttcat atcataaaag acttccattt
     4801 caatccaacc acattgagaa agtatcaaca ttttacagaa tccaatacac aagttgatat
     4861 agatcagcta cacagactaa tcaaccttgt atttggcaac ctcctcgctc acttccttga
     4921 gatcaccttt agtgaagatc aaaccaacgt ttccctaatc aaacaaacac agaaaaatcg
     4981 caaatttaca atctcaagtc tccagcgtat agaaagtctc aacattttcg aagaagctga
     5041 gattaacctg aaggagggga agcagattca aaattgcagt gtttccagag ttctcggagt
     5101 ggattctaac ggaacgtttc atcatagtgt tcttacccat gagaaccact gagtctccac
     5161 gaagaccttt cctgatgttc tgaagctgag tcgatccgac gttgtccgcc gcaacgacta
     5221 agatctgagt gtattcgtcg ataagctgac atagttttgt gtcatacgct atcttcttct
     5281 ctgcttttgt tgccttcacc attgttgctc aaaggattta gctttagata aaaactcaaa
     5341 atgttctctg tttcggagag tgtttgggaa agatgatgaa ggaagttagg gtttttgcgg
     5401 cgatctaaat ctaacggttc gtgtttgttt tgattaattt gtgatctgac ggctcttgtt
     5461 tactttgatg ggaattgggc tttaactaaa attaggccca aattgacaag ataacacgcg
     5521 tgacgttttt ttaatgttag aagaaaactt cgtgtcgttc attttggtaa aaactacaag
     5581 gcgttgatca ggagcttctt ctccttcttc ttcaccggga agagctcacc atgattctca
     5641 tcgttgtgga ttctctccga tcatctcgag aaagaggatc cacctgagga tcacaagttt
     5701 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     5761 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
     5821 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     5881 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     5941 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     6001 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     6061 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     6121 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     6181 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     6241 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     6301 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     6361 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     6421 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     6481 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
     6541 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc
     6601 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     6661 tatactgata tgtatacccg aagtatgtca aaaagaggtg tgctatgaag cagcgtatta
     6721 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     6781 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     6841 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     6901 tgctgacgag aacagggact ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     6961 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     7021 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     7081 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     7141 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     7201 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     7261 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     7321 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     7381 ttcttgtaca aagtggtgat cctccgttcc atgggctaga
//
