LOCUS       sequence-339200-        7865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 cloning vector for transient expression. Already contains 
            the 35S promoter and the Nos terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7865)
  AUTHORS   Grutzner R, Schubert R, Horn C, Yang C, Vogt T, Marillonnet S
  TITLE     Engineering Betalain Biosynthesis in Tomato for High Level Betanin 
            Production in Fruits.
  JOURNAL   Front Plant Sci. 2021 Jun 9;12:682443. doi: 
            10.3389/fpls.2021.682443. eCollection 2021.
  PUBMED    34177999
REFERENCE   2  (bases 1 to 7865)
  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: "Front Plant
            Sci."; date: "2021-06-9"; volume: "12:682443. doi"; pages: " 
            10.3389/fpls.2021.682443. eCollection 2021"
COMMENT     Sequence Label: pAGM53151
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      24..276
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1515..1539
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1695..2843)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3179..3767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3259..3278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3938..4729,4730..4798))
                     /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     4561..4580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4799..4903)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4990..5621
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5713..5737
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        6835..7180
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     7123..7143
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    7336..7357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7372..7377,7378..7395,7396..7402)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7410..7426
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7415..7437
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7434..7450
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7434..7450
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             7446..7769
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    7463..7519
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(7520..7537)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7520..7536)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7529..7551)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7745..7764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttgatcct ctagagtcaa gcagatcgtt caaacatttg gcaataaagt ttcttaagat
       61 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
      121 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
      181 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
      241 aattatcgcg cgcggtgtca tctatgttac tagatcgacg ctactattgt cttccactgg
      301 gtgtcaaagg agtgtcagtt aattaaagag tgtcggggcc gacactctta atggaagtaa
      361 aagtgtgggt tttgctgcac cgacactctt aatttaagag tgtcggggtc ccgatgaaac
      421 cgacgctttt aatttaagag tgtgggtttt tccacaccga cactcttatg aatgttaccc
      481 taaattcccc aatcctatac tgtataagag ctcattttta caacaattac caacaacaac
      541 aaacaacaaa caacattaca attacattta caattatcga tacaatgtct cctgaaagaa
      601 ggcctgctga tattagaagg gctactgagg ctgatatgcc tgctgtgtgc actattgtga
      661 atcattacat tgagacttct actgtgaatt tcaggactga gcctcaagaa ccacaagagt
      721 ggactgatga tcttgtgagg cttagagaga ggtatccttg gcttgttgct gaagtggatg
      781 gtgaagtggc tggtattgct tatgctggtc cttggaaggc taggaatgct tacgattgga
      841 ctgctgagtc tactgtttac gtgtctccta ggcatcaaag aactggtctt ggatctactc
      901 tttacactca tcttttgaag tctttggagg ctcagggttt caagtctgtg gttgctgtga
      961 ttggacttcc taatgatcct tctgtgagga tgcatgaggc tcttggttat gctcctaggg
     1021 gaatgcttag agctgctggt ttcaaacatg gaaattggca tgatgtggga ttctggcagc
     1081 ttgatttctc tcttcctgtt cctcctagac ctgttcttcc tgtgactgag atttgagctt
     1141 actccaccta aatactatgc cggttcaaat aagaaagtaa tgattatgta cccaaataat
     1201 acatttataa gatgtatgtg tgtgatggac atgtttctgt atcgtaccaa aactaaatag
     1261 gtggctttta tcccaaaaaa gtgctttcgt ttgatgacct tatgatttct aaagagtgtt
     1321 gttgttgctg cttatgttta ataaaagtga tgtatattat cagttgattt ccccacgtgg
     1381 aaggggtgat tatattgcac aaaattaagt taaaagtaac attgtggtgt aataaaaacc
     1441 ctagtgaatt tataacctat aagtaagtgg tgcagtagtg aatttatcac gaagtggttt
     1501 aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat
     1561 tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt
     1621 gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc agccggaact
     1681 gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca ggcgtgttcc
     1741 accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc
     1801 acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag cgggtacggc
     1861 tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac
     1921 ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg
     1981 atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc
     2041 agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt
     2101 aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga ccagcccagg
     2161 tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg
     2221 tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc gacgccggtg
     2281 taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc
     2341 gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct
     2401 cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc
     2461 tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc
     2521 tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac
     2581 ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc ggccgatggc
     2641 gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc
     2701 tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt
     2761 gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc
     2821 ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac tcacgccggg
     2881 gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag ataatccacc
     2941 ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta cttggtattc
     3001 cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc gtgggcctcg
     3061 gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt gtcgccggca
     3121 tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3181 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3241 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3301 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3361 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3421 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3481 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3541 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3601 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3661 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3721 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3781 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3841 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3901 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3961 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4021 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4081 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4141 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4201 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4261 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4321 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4381 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4441 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4501 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4561 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4621 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4681 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4741 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4801 tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg agcggataca
     4861 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat tggccagcgc
     4921 tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag
     4981 gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg
     5041 gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag
     5101 caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg
     5161 attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac
     5221 tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt
     5281 gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt
     5341 aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc
     5401 gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt
     5461 ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc
     5521 cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg
     5581 ccgccagccc agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag atcctgatct
     5641 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
     5701 ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa
     5761 taacacattg cggacgtttt taatgtactg gggtggatgc agtgggcccc actctgtgaa
     5821 gacaagcaag aattcaagct tggaggaatt ccaatcccac aaaaatctga gcttaacagc
     5881 acagttgctc ctctcagagc agaatcgggt attcaacacc ctcatatcaa ctactacgtt
     5941 gtgtataacg gtccacatgc cggtatatac gatgactggg gttgtacaaa ggcggcaaca
     6001 aacggcgttc ccggagttgc acacaagaaa tttgccacta ttacagaggc aagagcagca
     6061 gctgacgcgt acacaacaag tcagcaaaca gacaggttga acttcatccc caaaggagaa
     6121 gctcaactca agcccaagag ctttgctaag gccctaacaa gcccaccaaa gcaaaaagcc
     6181 cactggctca cgctaggaac caaaaggccc agcagtgatc cagccccaaa agagatctcc
     6241 tttgccccgg agattacaat ggacgatttc ctctatcttt acgatctagg aaggaagttc
     6301 gaaggtgaag gtgacgacac tatgttcacc actgataatg agaaggttag cctcttcaat
     6361 ttcagaaaga atgctgaccc acagatggtt agagaggcct acgcagcaag tctcatcaag
     6421 acgatctacc cgagtaacaa tctccaggag atcaaatacc ttcccaagaa ggttaaagat
     6481 gcagtcaaaa gattcaggac taattgcatc aagaacacag agaaagacat atttctcaag
     6541 atcagaagta ctattccagt atggacgatt caaggcttgc ttcataaacc aaggcaagta
     6601 atagagattg gagtctctaa aaaggtagtt cctactgaat ctaaggccat gcatggagtc
     6661 taagattcaa atcgaggatc taacagaact cgccgtcaag actggcgaac agttcataca
     6721 gagtctttta cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac
     6781 tctggtctac tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg ctattgagac
     6841 ttttcaacaa aggataattt cgggaaacct cctcggattc cattgcccag ctatctgtca
     6901 cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc attgcgataa
     6961 aggaaaggct atcattcaag atctctctgc cgacagtggt cccaaagatg gacccccacc
     7021 cacgaggagc atcgtggaaa aagaagaggt tccaaccacg tctacaaagc aagtggattg
     7081 atgtgacatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc
     7141 ttcctctata taaggaagtt catttcattt ggagaggaca cgctcgagta taagagctca
     7201 tttttacaac aattaccaac aacaacaaac aacaaacaac attacaatta catttacaat
     7261 tatcgataca atgtgagacc gcagctggca cgacaggttt cccgactgga aagcgggcag
     7321 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     7381 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7441 cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc
     7501 gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     7561 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     7621 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     7681 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     7741 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     7801 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgacgg
     7861 tctca
//
