LOCUS       sequence-438348-        7747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGGPK-AG2 where the ccdb/CmR has been swappe for mRFP1e; Esp3I 
            domesticated backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7747)
  AUTHORS   Pennetti VJ, LaFayette PR, Parrott WA
  TITLE     MultiGreen: A multiplexing architecture for GreenGate cloning.
  JOURNAL   PLoS One. 2024 Sep 18;19(9):e0306008. doi: 
            10.1371/journal.pone.0306008. eCollection 2024.
  PUBMED    39292669
REFERENCE   2  (bases 1 to 7747)
  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: "PLoS One.";
            date: "2024-09-18"; volume: "19(9):e0306008. doi"; pages: " 
            10.1371/journal.pone.0306008. eCollection 2024"
COMMENT     Sequence Label: pVP096
FEATURES             Location/Qualifiers
     source          1..7747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..97)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             89..904
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             89..904
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(162..181)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1500..2088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1989..2008
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2242..2259
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2274..2414
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2340..2359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2500..2522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2758..2952
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(3018..4091)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(4520..5149)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    6480..6504
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    6578..6599
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        6600..6634
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     RBS             6643..6654
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6661..7338
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     misc_feature    7367..7387
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     misc_feature    7519..7543
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tgtaacgatg acagagcgtt gctgcctgtg atccagatca tgaacaataa aactgtctgc
       61 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
      121 taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
      181 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
      241 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
      301 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
      361 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
      421 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
      481 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc ggctcgctca
      541 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
      601 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
      661 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
      721 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
      781 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
      841 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
      901 ctaacagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
      961 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     1021 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     1081 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     1141 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     1201 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta
     1261 gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg
     1321 cactaagcac ataattgctc acagccaaac tatcaggtca agtctgcttt tattattttt
     1381 aagcgtgcat aataagccct acacaaattg ggagatatat catgcatgac caaaatccct
     1441 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1501 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1561 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1621 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1681 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1741 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1801 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1861 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1921 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1981 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2041 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2101 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2161 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2221 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2281 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     2341 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     2401 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2461 tgctcccggc atccgcttac agacaagctg tgaccggctc cgggagctgc atgtgtcaga
     2521 ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc cttgatgtgg gcgccggcgg
     2581 tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg taggccagcc
     2641 atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg tagggagcgc
     2701 agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc agacagttat
     2761 gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta ggcggaaaaa
     2821 tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc aatgtacggc
     2881 tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt cccaatgtac
     2941 gtgctatcca caggaaagag agcttttcga cctttttccc ctgctagggc aatttgccct
     3001 agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag gttgcggtag
     3061 cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta ctccggcagg
     3121 tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc tccggcgctg
     3181 ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc tgcggcgcgg
     3241 cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta atcggggtga
     3301 accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca atcagctagc
     3361 tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta gcggctaatc
     3421 aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt cgtacgctgc
     3481 atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc tttctgcttt
     3541 ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg gaacacgcgg
     3601 ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag atgggaaacc
     3661 gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc tgcggaaacc
     3721 tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg gtcacgcttc
     3781 gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc cacgtcatag
     3841 agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct aatcgacggc
     3901 gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc cgcttgccac
     3961 gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg cgcggccttc
     4021 aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc ggatggtttg
     4081 cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag ggccggcaga
     4141 caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg cattccacgg
     4201 cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct aaaattcatc
     4261 tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc agatagcagc
     4321 tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc
     4381 ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc caacgttgca
     4441 gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct tttgctcatt
     4501 ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc agcgcctgga
     4561 cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg gcggcggcag
     4621 tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc tcgtacccgg
     4681 ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa
     4741 aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc tccaccaggt
     4801 cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg aagtcggctg
     4861 ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc actacgaagt
     4921 cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg
     4981 ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg ggatcggaat
     5041 cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg gttgcgatgg
     5101 tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg cgttcccctt
     5161 gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac gcaagctgtt
     5221 ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag cggccaagct
     5281 ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat gcagccgcac
     5341 ggttgatacg tgcgcgggcg gctcgaacac gtacccggcc gggatcatct ccgcctcgat
     5401 ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca tgcttgttcc
     5461 tcttggcgtt cattctcggc ggccgccagg gcgtcggcct cggtcaatgc gtcctcacgg
     5521 aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc aagtgcgcgg
     5581 tacagggtcg agcgatgcac gccaagcagt gcagccgcct ctttcacggt gcggccttcc
     5641 tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg gcgggggcca
     5701 aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc gatgattagg
     5761 gaacgctcga actcggcaat gccggcgaac acggtcaaca ccatgcggcc ggccggcgtg
     5821 gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg cgccggcctc ctggatgcgc
     5881 tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct ggtcactgtc
     5941 acaacgtcgc cagggcgtag gtggtcaagc atcctggcca gctccgggcg gtcgcgcctg
     6001 gtgccggtga tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag ttcggcccgt
     6061 tggttggtca agtcctggtc gtcggtgctg acgcgggcat agcccagcag gccagcggcg
     6121 gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc aagtattcta ctttatgcga
     6181 ctaaaacacg cgacaagaaa acgccaggaa aagggcaggg cggcagcctg tcgcgtaact
     6241 taggacttgt gcgacatgtc gttttcagaa gagggctgca ctgaacgtca gaagccgact
     6301 gcactatagc agcggagggg ttggatcaaa gtactttaaa gtactttaaa gtactttaaa
     6361 gtactttgat cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat
     6421 aaaccttttc acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg
     6481 tttacccgcc aatatatcct gtcaaacact gatagtttaa actgaaggcg ggaaacgaca
     6541 atctgatcca agctcaagct aagcttacct tgagaccgaa ttcgcggccg cttctagagt
     6601 ttacggctag ctcagtccta ggtacaatgc tagctactag agaaagagga gaaatactag
     6661 atggcttcct ccgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt
     6721 tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt
     6781 acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc
     6841 ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg
     6901 gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa
     6961 gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac
     7021 aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc
     7081 atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct gaaaggtgaa
     7141 atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc
     7201 tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac
     7261 atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt
     7321 cactccaccg gtgcttaata acgctgatag tgctagtgta gatcgctact agtagcggcc
     7381 gctgcaggtc tcagtatctg caggcatgcg acgtcgggcc ctctagagga tccccgggta
     7441 ccgcgaattc aattcggcgt taattcagta cattaaaaac gtccgcaatg tgttattaag
     7501 ttgtctaagc gtcaatttgt ttacaccaca atatatcctg ccaccagcca gccaacagct
     7561 ccccgaccgg cagctcggca caaaatcacc actcgataca ggcagcccat cagtccggga
     7621 cggcgtcagc gggagagccg ttgtaaggcg gcagactttg ctcatgttac cgatgctatt
     7681 cggaagaacg gcaactaagc tgccgggttt gaaacacgga tgatctcgcg gagggtagca
     7741 tgttgat
//
