LOCUS       sequence-316582-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tdTomato fluorescent protein-ER localization, 35 promoter, tNOS 
            terminator, transcriptional unit, inserted at the MoClo toolkit 
            level 1 acceptor position 1 reverse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  AUTHORS   Triozzi P, Schmidt HW, Dervinis C, Kirst M, Conde D
  TITLE     Simple, efficient and open-source CRISPR/Cas9 strategy for 
            multi-site genome editing in Populus tremula x alba.
  JOURNAL   Tree Physiol. 2021 May 7. pii: 6270869. doi: 
            10.1093/treephys/tpab066.
  PUBMED    33960379
REFERENCE   2  (bases 1 to 6992)
  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: "Tree 
            Physiol. 2021 May 7. pii: 6270869. doi: 10.1093/treephys/tpab066."
COMMENT     Sequence Label: pICH47802-p35S::tdTomato-ER::tNOS
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        409..754
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     697..717
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      2366..2618
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2729..2753
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      2845..3476
                     /label=oriV
                     /note="incP origin of replication"
     promoter        3563..3667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3668..3736,3737..4528)
                     /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(3886..3905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4699..5287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5188..5207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5623..6771
                     /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    6927..6951
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggaggtcaac atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt
       61 ctcagaagat caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct
      121 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg
      181 ctcctacaaa tgccatcatt gcgataaagg aaaggctatc attcaagatc tctctgccga
      241 cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagaggttcc
      301 aaccacgtct acaaagcaag tggattgatg tgataacatg gtggagcacg acactctggt
      361 ctactccaaa aatgtcaaag atacagtctc agaagatcaa agggctattg agacttttca
      421 acaaaggata atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
      481 cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa
      541 ggctatcatt caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag
      601 gagcatcgtg gaaaaagaag aggttccaac cacgtctaca aagcaagtgg attgatgtga
      661 catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
      721 tatataagga agttcatttc atttggagag gacacgctcg agtataagag ctcattttta
      781 caacaattac caacaacaac aaacaacaaa caacattaca attacattta caattatcga
      841 tacaatggtc aagactaatc ttttcctttt cttcatcttc tcgcttctcc tctcactttc
      901 cagcgcggac atggtgtcca agggcgaaga ggtgattaaa gagttcatga ggttcaaagt
      961 gagaatggag ggcagtatga acggccatga gttcgagatc gaaggtgaag gcgaaggtag
     1021 accatacgag ggcacccaga cggcgaaact gaaagtgacg aagggcggcc cactgccctt
     1081 cgcctgggac attctgagtc cacaatttat gtacggttcc aaggcatacg ttaagcaccc
     1141 cgccgacatc cctgattaca aaaagctgtc gttcccggag gggtttaaat gggaaagagt
     1201 gatgaatttt gaggacggag ggctcgtcac tgtgactcaa gactcctcac ttcaagacgg
     1261 caccctaatt tacaaggtga agatgagggg aacaaatttc ccccctgatg gccctgttat
     1321 gcagaaaaaa acaatgggat gggaggcatc caccgagaga ctttacccgc gcgacggagt
     1381 gctcaaaggc gaaatccatc aggcactgaa actaaaggac ggcggccact acctggttga
     1441 gttcaaaact atatatatgg ccaagaaacc cgtccagttg ccgggatatt actatgtgga
     1501 cacaaaactg gatataacgt cccacaacga agattacacc atagtggaac agtatgagcg
     1561 cagtgagggt cgtcaccatc tattcctcgg acatggcacg ggttccacgg gctcgggttc
     1621 cagcggcacg gcaagctcgg aggacaacaa catggcggtg atcaaagagt tcatgcggtt
     1681 taaagtccgt atggagggca gcatgaatgg gcacgaattt gaaattgagg gagaaggcga
     1741 gggccgacca tatgagggga cacaaaccgc gaaactaaaa gtcaccaaag gtgggccgct
     1801 tccttttgcc tgggacattc tctctcctca gttcatgtat ggctccaagg cctacgttaa
     1861 acatccggct gatatccccg attataagaa attgtctttc ccggaggggt ttaaatggga
     1921 gcgggttatg aatttcgagg atggtggact ggttactgtt acacaagatt cgagtctgca
     1981 ggatggcaca ttgatataca aagtcaaaat gaggggtacg aattttcccc cggacggccc
     2041 ggtgatgcag aaaaaaacta tgggctggga ggccagcacg gagaggctgt acccgaggga
     2101 cggggttttg aaaggggaga tacaccaggc gcttaaactg aaggacgggg ggcactatct
     2161 ggtggaattt aagacgatat atatggcgaa aaagcccgtg cagctacctg gctactacta
     2221 cgtcgatacc aaactcgata ttacctctca taacgaggac tacactatcg tggagcaata
     2281 tgaaagatca gaaggacgtc accacctttt tctatacgga atggacgagc tttacaaaaa
     2341 ggacgagttg taagcttgtc aagcagatcg ttcaaacatt tggcaataaa gtttcttaag
     2401 attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa
     2461 gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag
     2521 agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga
     2581 taaattatcg cgcgcggtgt catctatgtt actagatcga cgctggatcc gaattcggca
     2641 ttgtcttcac agagtggggc ccactgcatc caccccagta cattaaaaac gtccgcaatg
     2701 tgttattaag ttgtctaagc gtcaatttgt ttacaccaca atatatcctg ccaccagcca
     2761 gccaacagct ccccgaccgg cagctcggca caaaatcacc actcgataca ggcagcccat
     2821 cagtcagatc aggatctcct ttgcgacgct caccgggctg gttgccctcg ccgctgggct
     2881 ggcggccgtc tatggccctg caaacgcgcc agaaacgccg tcgaagccgt gtgcgagaca
     2941 ccgcggccgc cggcgttgtg gatacctcgc ggaaaacttg gccctcactg acagatgagg
     3001 ggcggacgtt gacacttgag gggccgactc acccggcgcg gcgttgacag atgaggggca
     3061 ggctcgattt cggccggcga cgtggagctg gccagcctcg caaatcggcg aaaacgcctg
     3121 attttacgcg agtttcccac agatgatgtg gacaagcctg gggataagtg ccctgcggta
     3181 ttgacacttg aggggcgcga ctactgacag atgaggggcg cgatccttga cacttgaggg
     3241 gcagagtgct gacagatgag gggcgcacct attgacattt gaggggctgt ccacaggcag
     3301 aaaatccagc atttgcaagg gtttccgccc gtttttcggc caccgctaac ctgtctttta
     3361 acctgctttt aaaccaatat ttataaacct tgtttttaac cagggctgcg ccctgtgcgc
     3421 gtgaccgcgc acgccgaagg ggggtgcccc cccttctcga accctcccgg cccgctaacg
     3481 cgggcctccc atccccccag gggctgcgcc cctcggccgc gaacggcctc accccaaaaa
     3541 tggcagcgct ggccaattcg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3601 aaatatgtat ccgctcatga gacaataacc ttgataaatg cttcaataat attgaaaaag
     3661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4381 tggttctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4681 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4801 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5101 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5281 atggaaaaac gccagcaacg cggccttttt acggttcctg gcagatccta gatgtggcgc
     5341 aacgatgccg gcgacaagca ggagcgcacc gacttcttcc gcatcaagtg ttttggctct
     5401 caggccgagg cccacggcaa gtatttgggc aaggggtcgc tggtattcgt gcagggcaag
     5461 attcggaata ccaagtacga gaaggacggc cagacggtct acgggaccga cttcattgcc
     5521 gataaggtgg attatctgga caccaaggca ccaggcgggt caaatcagga ataagggcac
     5581 attgccccgg cgtgagtcgg ggcaatcccg caaggagggt gaatgaatcg gacgtttgac
     5641 cggaaggcat acaggcaaga actgatcgac gcggggtttt ccgccgagga tgccgaaacc
     5701 atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct tccagtccgt cggctcgatg
     5761 gtccagcaag ctacggccaa gatcgagcgc gacagcgtgc aactggctcc ccctgccctg
     5821 cccgcgccat cggccgccgt ggagcgttcg cgtcgtcttg aacaggaggc ggcaggtttg
     5881 gcgaagtcga tgaccatcga cacgcgagga actatgacga ccaagaagcg aaaaaccgcc
     5941 ggcgaggacc tggcaaaaca ggtcagcgag gccaagcagg ccgcgttgct gaaacacacg
     6001 aagcagcaga tcaaggaaat gcagctttcc ttgttcgata ttgcgccgtg gccggacacg
     6061 atgcgagcga tgccaaacga cacggcccgc tctgccctgt tcaccacgcg caacaagaaa
     6121 atcccgcgcg aggcgctgca aaacaaggtc attttccacg tcaacaagga cgtgaagatc
     6181 acctacaccg gcgtcgagct gcgggccgac gatgacgaac tggtgtggca gcaggtgttg
     6241 gagtacgcga agcgcacccc tatcggcgag ccgatcacct tcacgttcta cgagctttgc
     6301 caggacctgg gctggtcgat caatggccgg tattacacga aggccgagga atgcctgtcg
     6361 cgcctacagg cgacggcgat gggcttcacg tccgaccgcg ttgggcacct ggaatcggtg
     6421 tcgctgctgc accgcttccg cgtcctggac cgtggcaaga aaacgtcccg ttgccaggtc
     6481 ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc actacacgaa attcatatgg
     6541 gagaagtacc gcaagctgtc gccgacggcc cgacggatgt tcgactattt cagctcgcac
     6601 cgggagccgt acccgctcaa gctggaaacc ttccgcctca tgtgcggatc ggattccacc
     6661 cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg aagagttgcg aggcagcggc
     6721 ctggtggaac acgcctgggt caatgatgac ctggtgcatt gcaaacgcta gggccttgtg
     6781 gggtcagttc cggctggggg ttcagcagcc agcgcctgat ctggggaacc ctgtggttgg
     6841 catgcacata caaatggacg aacggataaa ccttttcacg cccttttaaa tatccgatta
     6901 ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc aaacactgat
     6961 agtttaaacc acttcgtgca gaagacaatt gc
//
