LOCUS       sequence-312949-        6509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1, Position 2 Golden Gate vector. HspHv17::Cre-U5-Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6509)
  AUTHORS   Harrington SA, Backhaus AE, Fox S, Rogers C, Borrill P, Uauy C, 
            Richardson A
  TITLE     A heat-shock inducible system for flexible gene expression in 
            cereals.
  JOURNAL   Plant Methods. 2020 Oct 14;16:137. doi: 10.1186/s13007-020-00677-3. 
            eCollection 2020.
  PUBMED    33072173
REFERENCE   2  (bases 1 to 6509)
  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: "Plant 
            Methods."; date: "2020-10-14"; volume: "16:137. doi"; pages: " 
            10.1186/s13007-020-00677-3. eCollection 2020"
COMMENT     Sequence Label: EC71173
FEATURES             Location/Qualifiers
     source          1..6509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    116..140
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      232..863
                     /label=oriV
                     /note="incP origin of replication"
     promoter        950..1054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1055..1123,1124..1915)
                     /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(1273..1292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2086..2674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2575..2594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3010..4158
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4314..4338
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     intron          5279..5467
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     terminator      join(6268..6509,1..7)
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
ORIGIN
        1 agataagcgc taagcttgaa ttcttgcttg tcttcacaga gtggggccca ctgcatccac
       61 cccagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc aatttgttta
      121 caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag ctcggcacaa
      181 aatcaccact cgatacaggc agcccatcag tcagatcagg atctcctttg cgacgctcac
      241 cgggctggtt gccctcgccg ctgggctggc ggccgtctat ggccctgcaa acgcgccaga
      301 aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg cgttgtggat acctcgcgga
      361 aaacttggcc ctcactgaca gatgaggggc ggacgttgac acttgagggg ccgactcacc
      421 cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg ccggcgacgt ggagctggcc
      481 agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt ttcccacaga tgatgtggac
      541 aagcctgggg ataagtgccc tgcggtattg acacttgagg ggcgcgacta ctgacagatg
      601 aggggcgcga tccttgacac ttgaggggca gagtgctgac agatgagggg cgcacctatt
      661 gacatttgag gggctgtcca caggcagaaa atccagcatt tgcaagggtt tccgcccgtt
      721 tttcggccac cgctaacctg tcttttaacc tgcttttaaa ccaatattta taaaccttgt
      781 ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg ccgaaggggg gtgccccccc
      841 ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc cccccagggg ctgcgcccct
      901 cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc caattcgtgc gcggaacccc
      961 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccttg
     1021 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1081 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1141 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1201 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1261 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1321 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1381 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1441 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1501 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1561 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1621 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1681 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1741 tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt atcattgcag cactggggcc
     1801 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1861 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1921 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1981 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2041 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2101 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2161 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2221 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2281 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2341 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2401 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2461 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2521 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2581 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2641 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2701 gttcctggca gatcctagat gtggcgcaac gatgccggcg acaagcagga gcgcaccgac
     2761 ttcttccgca tcaagtgttt tggctctcag gccgaggccc acggcaagta tttgggcaag
     2821 gggtcgctgg tattcgtgca gggcaagatt cggaatacca agtacgagaa ggacggccag
     2881 acggtctacg ggaccgactt cattgccgat aaggtggatt atctggacac caaggcacca
     2941 ggcgggtcaa atcaggaata agggcacatt gccccggcgt gagtcggggc aatcccgcaa
     3001 ggagggtgaa tgaatcggac gtttgaccgg aaggcataca ggcaagaact gatcgacgcg
     3061 gggttttccg ccgaggatgc cgaaaccatc gcaagccgca ccgtcatgcg tgcgccccgc
     3121 gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta cggccaagat cgagcgcgac
     3181 agcgtgcaac tggctccccc tgccctgccc gcgccatcgg ccgccgtgga gcgttcgcgt
     3241 cgtcttgaac aggaggcggc aggtttggcg aagtcgatga ccatcgacac gcgaggaact
     3301 atgacgacca agaagcgaaa aaccgccggc gaggacctgg caaaacaggt cagcgaggcc
     3361 aagcaggccg cgttgctgaa acacacgaag cagcagatca aggaaatgca gctttccttg
     3421 ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc caaacgacac ggcccgctct
     3481 gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg cgctgcaaaa caaggtcatt
     3541 ttccacgtca acaaggacgt gaagatcacc tacaccggcg tcgagctgcg ggccgacgat
     3601 gacgaactgg tgtggcagca ggtgttggag tacgcgaagc gcacccctat cggcgagccg
     3661 atcaccttca cgttctacga gctttgccag gacctgggct ggtcgatcaa tggccggtat
     3721 tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga cggcgatggg cttcacgtcc
     3781 gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc gcttccgcgt cctggaccgt
     3841 ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg aaatcgtcgt gctgtttgct
     3901 ggcgaccact acacgaaatt catatgggag aagtaccgca agctgtcgcc gacggcccga
     3961 cggatgttcg actatttcag ctcgcaccgg gagccgtacc cgctcaagct ggaaaccttc
     4021 cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt ggcgcgagca ggtcggcgaa
     4081 gcctgcgaag agttgcgagg cagcggcctg gtggaacacg cctgggtcaa tgatgacctg
     4141 gtgcattgca aacgctaggg ccttgtgggg tcagttccgg ctgggggttc agcagccagc
     4201 gcctgatctg gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct
     4261 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
     4321 cgccaatata tcctgtcaaa cactgatagt ttaaaccact tcgtgcagaa gacaatagtg
     4381 gagaatttta agatggtgca tggaccaccc ggaccacccg ctagctgcgt cattgtgtag
     4441 tactctttct gttttaaaat aagtgtctga actttatact aactttaata taaaattgta
     4501 ctaaacttga gacatttatt ttaggacgga gggagtagta tttatttacg tttaactatt
     4561 gtacttgctc gctgtaactt taatgacaga taacctgtac tgttaatcat gctattctgg
     4621 aaacggcccc cacaacccat tcttgaatct agactagtcg cccccttgct ggaattatac
     4681 aattatttat agaaaaatct cacaaaaaaa ttattaacaa aaaaaaaagg gagaaggcgc
     4741 cctaacggca ggttcgtgag tcccctgcaa gacgatagac ccttcgaggg ttgggaacag
     4801 gaaagaacga atggctcaca gcctacctgg aaagcgaaag agaaacaaac actcgtcgat
     4861 tggacagtcc tcgagattct tcaagatcag tacctacgat gctataagta gtggctcacc
     4921 ttccatcgcc ctccatccca actcaagcca gcgtcaagca aaagcgaaca acatcctacc
     4981 tacaatccga tacagattaa tttcaacgcg agctacaaac caaagcaagc aacaccgacc
     5041 aatgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag
     5101 tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca
     5161 tacctggaaa atgctcctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa
     5221 ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggt
     5281 aagtttctgc ttctaccttt gatatatata taataattat cattaattag tagtaatata
     5341 atatttcaaa tatttttttc aaaataaaag aatgtggtat atagcaattg cttttctgta
     5401 gtttataagt gtgtatattt taatttataa cttttctaat atatgaccaa aatttgttga
     5461 tgtgcaggcg cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat
     5521 gcttcatcgt cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat
     5581 gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt
     5641 cgaacgcact gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga
     5701 tatacgtaat ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat
     5761 tgccaggatc agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat
     5821 tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag aaggcactta gcctgggggt
     5881 aactaaactg gtcgagcgat ggatttccgt ttctggtgta gctgatgatc cgaataacta
     5941 cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca gccagctatc
     6001 aactcgcgcc ctggaaggga tttttgaagc aactcatcga ttgatttacg gcgctaagga
     6061 tgactctggt cagagatacc tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg
     6121 agatatggcc cgcgctggag tttcaatacc ggagatcatg caagctggtg gctggaccaa
     6181 tgtaaatatt gtcatgaact atatccgtaa cctggatagt gaaacagggg caatggtgcg
     6241 cctgctggaa gatggcgatt aggcttatat gaagatgaag atgaaatatt tggtgtgtca
     6301 aataaaaagc ttgtgtgctt aagtttgtgt ttttttcttg gcttgttgtg ttatgaattt
     6361 gtggcttttt ctaatattaa atgaatgtaa gatctcatta taatgaataa acaaatgttt
     6421 ctataatcca ttgtgaatgt tttgttggat ctcttctgca gcatataact actgtatgtg
     6481 ctatggtatg gactatggaa tatgattaa
//
