LOCUS       sequence-492313-       10493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pL1-R2-pAtUBI-RUBY-tRBCS (Golden Gate Level 1 module with a 
            transcription unit expressing RUBY from the AtUBI promoter with the 
            tRBC terminator for R2 position in Level 2 vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10493)
  TITLE     Monitoring the stability of transgene expression in lettuce using 
            the RUBY reporter.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10493)
  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            "Unpublished"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: pR2B5
FEATURES             Location/Qualifiers
     source          1..10493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1526..5476
                     /label=RUBY
     CDS             3026..3082
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3917..3973
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     terminator      5482..6123
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    6236..6260
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      6352..6983
                     /label=oriV
                     /note="incP origin of replication"
     promoter        7070..7174
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7175..7243,7244..8035)
                     /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(7393..7412)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8206..8794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8695..8714
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             9130..10278
                     /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    10434..10458
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 aatagtggag gacgtcgttg tggttggtgc tttccttaca ttctgagcct ctttccttct
       61 aatccactca tctgcatctt cttgtgtcct tactaatacc tcattggttc caaattccct
      121 ccctttaagc accagctcgt ttctgttctt ccacagcctc ccaagtatcc aagggactaa
      181 agcctccaca ttcttcagat caggatattc ttgtttaaga tgttgaactc tatggaggtt
      241 tgtatgaact gatgatctag gaccggataa gttcccttct tcatagcgaa cttattcaaa
      301 gaatgttttg tgtatcattc ttgttacatt gttattaatg aaaaaatatt attggtcatt
      361 ggactgaaca cgagtgttaa atatggacca ggccccaaat aagatccatt gatatatgaa
      421 ttaaataaca agaataaatc gagtcaccaa accacttgcc ttttttaacg agacttgttc
      481 accaacttga tacaaaagtc attatcctat gcaaatcaat aatcatacaa aaatatccaa
      541 taacactaaa aaaattaaaa gaaatggata atttcacaat atgttatacg ataaagaagt
      601 tacttttcca agaaattcac tgattttata agcccacttg cattagataa atggcaaaaa
      661 aaaacaaaaa ggaaaagaaa taaagcacga agaattctag aaaatacgaa atacgcttca
      721 atgcagtggg acccacggtt caattattgc caattttcag ctccaccgta tatttaaaaa
      781 ataaaacgat aatgctaaaa aaatataaat cgtaacgatc gttaaatctc aacggctgga
      841 tcttatgacg accgttaagg aaattgtggt tgtcggacga agtccagtaa taaacggcgt
      901 caaagtggtt gcagccggca cacacgagtc gtgtttatca actcaaagca caaatacttt
      961 tcctcaacct aaaaataagg caattagcca aaaacaactt tgcgtgtaaa caacgctcaa
     1021 tacacgtgtc attttattat tagctattgc ttcaccgcct tagctttctc gtgacctagt
     1081 cgtcctcgtc ttttcttctt cttcttctat aaaacaatac ccaaagagct cttcttcttc
     1141 acaattcaga tttcaatttc tcaaaatctt aaaaactttc tctcaattct ctctaccgtg
     1201 atcaaggtaa atttctgtgt tccttattct ctcaaaatct tcgattttgt tttcgttcga
     1261 tcccaatttc gtatatgttc tttggtttag attctgttaa tcttagatcg tagacgattt
     1321 tctgggtttg atcgttagat atcatcttaa ttctcgatta gggtttcata gatatcatcc
     1381 gatttgttca aataatttga gttttgtcga ataattactc ttcgatttgt gatttctatc
     1441 tagatctggt gttagtttct agtttgtgcg atcgaatttg tcgattaatc tgagtttttc
     1501 tgattaacag atgcagatct taatgatgga tcatgcgacc ctcgccatga tcctcgcgat
     1561 ctggttcatc agcttccact tcatcaagct gctgttctcc cagcagacca ccaagctgct
     1621 tccgccagga ccaaagccgc ttccgatcat cggcaacatc cttgaggtgg gcaagaagcc
     1681 gcatcggtcc ttcgccaacc tcgccaagat tcacggccca ctcatttccc tcagactcgg
     1741 ctctgtgacc accatcgttg tgtcctctgc cgacgtggcc aaagagatgt tcctcaagaa
     1801 ggatcacccg ctctccaacc gcacgatccc gaatagtgtt acagccggcg accaccacaa
     1861 gctcaccatg tcttggctcc cggtgtctcc gaagtggcgc aacttccgca agattaccgc
     1921 cgtgcatctg ctctccccac agagactcga tgcctgccag acattcaggc acgccaaggt
     1981 gcagcagctc tacgagtacg ttcaagagtg cgcccagaaa ggccaggccg tggatattgg
     2041 caaggccgcc tttacgacca gcctcaacct cctcagcaag ctgttcttca gcgtcgagct
     2101 ggcgcaccac aagtcccata ccagccaaga gttcaaagag ctgatctgga acatcatgga
     2161 agatataggc aagccgaact acgccgacta cttcccgatt ctcggctgcg ttgacccatc
     2221 tggcattaga agaaggctcg cctgctcctt cgacaagctg atcgccgtgt tccagggcat
     2281 catctgcgag agactcgccc cagattcctc caccacaact accaccacca ccgacgacgt
     2341 gctcgatgtg ctcctccagc tgttcaagca gaacgagctg acgatgggcg agatcaacca
     2401 cctcctcgtg gacatcttcg acgccggcac cgataccaca tcctccacat tcgagtgggt
     2461 gatgaccgag ctgatccgca atccagagat gatggaaaag gcccaagagg aaatcaagca
     2521 ggtcctcggc aaggacaagc agatccaaga gtccgacatc atcaacctgc cgtacctcca
     2581 ggcgatcatc aaagagacac tccgcctcca tccgccgacc gtgttcttgc tcccaagaaa
     2641 ggccgacacc gatgtcgagc tgtacggcta catcgtgccg aaggatgccc agatcctcgt
     2701 gaacctctgg gccattggca gggacccaaa cgcctggcag aacgccgata ttttcagccc
     2761 agagcgcttc atcggctgcg agatcgatgt taagggccgc gatttcggcc tccttccatt
     2821 tggcgctggc cgcagaattt gcccaggcat gaatctcgcc atcaggatgc tcaccctcat
     2881 gctcgccaca ctcctccagt tcttcaactg gaagctcgaa ggcgacatct ccccgaagga
     2941 cctcgacatg gacgagaagt tcggcattgc gctccaaaag accaagccgc tcaagctcat
     3001 cccgattccg cgctacggta gcggagctac caattttagc ctccttaagc aggcaggtga
     3061 tgtagaagag aaccccgggc ctatgaagat gatgaacggc gaggacgcca acgaccagat
     3121 gatcaaagag tccttcttca tcacccacgg caacccgatc ctcaccgtcg aggatacaca
     3181 tccgctcagg ccgttcttcg agacatggcg cgagaagatt ttctccaaga agccgaaggc
     3241 catcctcatc atctccggcc actgggagac agtgaagcca accgtgaacg ccgtgcacat
     3301 caacgacacc atccacgact tcgacgacta cccagccgcc atgtaccagt tcaagtaccc
     3361 agctccaggc gagccagagc ttgcgagaaa ggtggaagag atcctcaaga agtccgggtt
     3421 cgagacagcc gagacagacc aaaagagggg ccttgatcac ggcgcctggg ttccactcat
     3481 gctcatgtat ccagaggcgg acatcccggt gtgccagctc tcagttcagc cacatctcga
     3541 cggcacctac cactacaatc tcggcagagc cctcgcgccg ctcaagaatg atggcgtgct
     3601 cattattggc tccggcagcg ccacacatcc actcgatgag acaccgcact acttcgatgg
     3661 tgttgcccct tgggccgctg ccttcgattc ttggcttagg aaggccctca tcaacggccg
     3721 cttcgaggaa gtgaacatct acgagagcaa ggccccgaac tggaagctcg cccatccatt
     3781 tccagagcac ttctacccgc tccacgttgt gctcggcgct gctggtgaaa agtggaaggc
     3841 cgagctgatc cactcctcct gggatcatgg cacactttgc cacggctcct acaagttcac
     3901 ctccgccgga tccggagcaa ccaactttag cctgctcaag caagcaggag atgttgagga
     3961 aaatcctggc cccatgaccg ccatcaagat gaacaccaac ggcgagggcg agacacagca
     4021 catcctcatg atcccgttca tggcgcaggg ccacctcagg ccatttctcg aactcgccat
     4081 gttcctctac aagcgctccc acgtgatcat caccctgctc acaactccgc tcaatgccgg
     4141 cttcctcagg cacctccttc accaccattc ctactcctcc agcggcatca ggatcgtcga
     4201 gctgccattc aactccacca accacggact cccaccgggc atcgagaaca ccgataagct
     4261 cacactcccg ctcgtggtgt ccctcttcca ttccaccatc agcctcgatc cgcacctccg
     4321 cgattacatc tccaggcatt tcagcccagc caggccacca ctctgcgtga tccatgatgt
     4381 gttcctcggc tgggttgacc aggtggccaa ggatgtgggc tctacaggcg tggtgttcac
     4441 aacaggcggc gcttatggca catccgccta cgtgtccatc tggaacgatc tcccgcacca
     4501 gaactactcc gacgaccaag agttcccgct gccaggcttc ccagagaacc ataagttccg
     4561 caggtcccag ctccatcggt tcctcagata tgccgacggc tccgacgatt ggtccaagta
     4621 tttccagccg cagctccgcc agtccatgaa gtcttttggc tggctctgca actccgtgga
     4681 agagatcgag acactcggct tctccatcct ccgcaactac accaagctgc cgatctgggg
     4741 catcggccca cttattgctt ccccagtgca gcactcctcc tccgacaaca attcaacagg
     4801 cgccgagttc gtgcagtggc tcagcctcaa agagccggac tccgtcctct acatctcctt
     4861 cggctcccag aacacgatca gcccgacgca gatgatggaa ctcgctgctg gccttgagtc
     4921 ctccgagaag ccattcctct gggtgatcag agccccgttc ggcttcgaca tcaacgaaga
     4981 gatgcgccca gagtggctgc cagagggctt tgaggaacgc atgaaggtga agaaacaggg
     5041 caagctcgtg tacaagctcg gcccgcagct tgagatcctc aaccatgaat ccatcggcgg
     5101 ctttctcacc cactgcggat ggaacagcat ccttgagtct cttcgcgagg gcgttccgat
     5161 gcttggatgg ccacttgctg ccgagcaggc ctacaacctc aagtacctcg aagatgagat
     5221 gggcgtcgcg gttgagcttg ctagaggcct cgaaggcgag atctccaaag agaaggtcaa
     5281 gcgcatcgtc gagatgatcc ttgagcgcaa cgagggctcc aaaggctggg agatgaagaa
     5341 tcgcgccgtg gaaatgggca aaaagctcaa ggacgccgtg aacgaggaaa aagagctgaa
     5401 gggctcctcc gtgaaggcga tcgacgattt cctcgacgcc gtcatgcagg ccaaacttga
     5461 gccaagcctc cagtgagctt gagctttcgt tcgtatcatc ggtttcgaca acgttcgtca
     5521 agttcaatgc atcagtttca ttgcgcacac accagaatcc tactgagttc gagtattatg
     5581 gcattgggaa acatgttttt cttgtaccat ttgttgtgct tgtaatttac tgtgtttttt
     5641 attcggtttt cgctatcgaa ctgtgaaatg gaaatggatg gagaagagtt aatgaatgat
     5701 atggtccttt tgttcattct caaattaata ttatttgttt tttctcttat ttgttgtgtg
     5761 ttgaatttga aaatataaga gatatgcaaa cattttgttt tgagtaaaaa tgtgtcaaat
     5821 cgtggcctct aatgaccgaa gttaatatga ggagtaaaac acttgtagtt gtaccattat
     5881 gcttattcac taggcaacaa atatattttc agacctagaa aagctgcaaa tgttactgaa
     5941 tacaagtatg tcctcttgtg ttttagacat ttatgaactt tcctttatgt aattttccag
     6001 aatccttgtc agattctaat cattgcttta taattatagt tatactcatg gatttgtagt
     6061 tgagtatgaa aatatttttt aatgcatttt atgacttgcc aattgattga caacatgcat
     6121 caatcgacgc taagcttgaa ttcttgcttg tcttcacaga gtggggccca ctgcatccac
     6181 cccagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc aatttgttta
     6241 caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag ctcggcacaa
     6301 aatcaccact cgatacaggc agcccatcag tcagatcagg atctcctttg cgacgctcac
     6361 cgggctggtt gccctcgccg ctgggctggc ggccgtctat ggccctgcaa acgcgccaga
     6421 aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg cgttgtggat acctcgcgga
     6481 aaacttggcc ctcactgaca gatgaggggc ggacgttgac acttgagggg ccgactcacc
     6541 cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg ccggcgacgt ggagctggcc
     6601 agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt ttcccacaga tgatgtggac
     6661 aagcctgggg ataagtgccc tgcggtattg acacttgagg ggcgcgacta ctgacagatg
     6721 aggggcgcga tccttgacac ttgaggggca gagtgctgac agatgagggg cgcacctatt
     6781 gacatttgag gggctgtcca caggcagaaa atccagcatt tgcaagggtt tccgcccgtt
     6841 tttcggccac cgctaacctg tcttttaacc tgcttttaaa ccaatattta taaaccttgt
     6901 ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg ccgaaggggg gtgccccccc
     6961 ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc cccccagggg ctgcgcccct
     7021 cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc caattcgtgc gcggaacccc
     7081 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccttg
     7141 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7201 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7261 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7321 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7381 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7441 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7501 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7561 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7621 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7681 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7741 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7801 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7861 tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt atcattgcag cactggggcc
     7921 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7981 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     8041 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8101 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8161 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     8221 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8281 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8341 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8401 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     8461 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     8521 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     8581 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     8641 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8701 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8761 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8821 gttcctggca gatcctagat gtggcgcaac gatgccggcg acaagcagga gcgcaccgac
     8881 ttcttccgca tcaagtgttt tggctctcag gccgaggccc acggcaagta tttgggcaag
     8941 gggtcgctgg tattcgtgca gggcaagatt cggaatacca agtacgagaa ggacggccag
     9001 acggtctacg ggaccgactt cattgccgat aaggtggatt atctggacac caaggcacca
     9061 ggcgggtcaa atcaggaata agggcacatt gccccggcgt gagtcggggc aatcccgcaa
     9121 ggagggtgaa tgaatcggac gtttgaccgg aaggcataca ggcaagaact gatcgacgcg
     9181 gggttttccg ccgaggatgc cgaaaccatc gcaagccgca ccgtcatgcg tgcgccccgc
     9241 gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta cggccaagat cgagcgcgac
     9301 agcgtgcaac tggctccccc tgccctgccc gcgccatcgg ccgccgtgga gcgttcgcgt
     9361 cgtcttgaac aggaggcggc aggtttggcg aagtcgatga ccatcgacac gcgaggaact
     9421 atgacgacca agaagcgaaa aaccgccggc gaggacctgg caaaacaggt cagcgaggcc
     9481 aagcaggccg cgttgctgaa acacacgaag cagcagatca aggaaatgca gctttccttg
     9541 ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc caaacgacac ggcccgctct
     9601 gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg cgctgcaaaa caaggtcatt
     9661 ttccacgtca acaaggacgt gaagatcacc tacaccggcg tcgagctgcg ggccgacgat
     9721 gacgaactgg tgtggcagca ggtgttggag tacgcgaagc gcacccctat cggcgagccg
     9781 atcaccttca cgttctacga gctttgccag gacctgggct ggtcgatcaa tggccggtat
     9841 tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga cggcgatggg cttcacgtcc
     9901 gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc gcttccgcgt cctggaccgt
     9961 ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg aaatcgtcgt gctgtttgct
    10021 ggcgaccact acacgaaatt catatgggag aagtaccgca agctgtcgcc gacggcccga
    10081 cggatgttcg actatttcag ctcgcaccgg gagccgtacc cgctcaagct ggaaaccttc
    10141 cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt ggcgcgagca ggtcggcgaa
    10201 gcctgcgaag agttgcgagg cagcggcctg gtggaacacg cctgggtcaa tgatgacctg
    10261 gtgcattgca aacgctaggg ccttgtgggg tcagttccgg ctgggggttc agcagccagc
    10321 gcctgatctg gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct
    10381 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
    10441 cgccaatata tcctgtcaaa cactgatagt ttaaaccact tcgtgcagaa gac
//
