LOCUS       sequence-258710-        3435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of mCerulean3 for C-term fusions in the 
            pGGD000 backbone (GreenGate).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3435)
  AUTHORS   Denay G, Schultz P, Hansch S, Weidtkamp-Peters S, Simon R
  TITLE     Over the rainbow: A practical guide for fluorescent protein 
            selection in plant FRET experiments.
  JOURNAL   Plant Direct. 2019 Dec 6;3(12):e00189. doi: 10.1002/pld3.189. 
            eCollection 2019 Dec.
  PUBMED    31844834
REFERENCE   2  (bases 1 to 3435)
  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 
            Direct."; date: "2019-12-6"; volume: "3(12):e00189. doi"; pages: " 
            10.1002/pld3.189. eCollection 2019 Dec"
COMMENT     Sequence Label: pGD251
FEATURES             Location/Qualifiers
     source          1..3435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(15..20,21..38,39..45)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    53..69
                     /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)."
     CDS             join(152..154,155..157,158..871)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(197..218)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(458..477)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     805..826
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(892..911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(893..911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(921..943)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1137..1156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1256..1278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1316..1334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1402..1506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1507..1575,1576..2367)
                     /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(1725..1744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2538..3126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3027..3046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3280..3297
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3414..3435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
       61 ggataacaat ttcacacgag ctatttaggt gacattatag aagtgaagct tggtctcatc
      121 aggcagcggc tctggatcgg cggccgctgc aatggtgagc aagggcgagg agctgttcac
      181 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      241 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      301 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgagct ggggcgtgca
      361 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      421 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      481 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      541 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacgcca tccacggcaa
      601 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttcg gcctcaactg
      661 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      721 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
      781 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      841 cactctcggc atggacgagc tgtacaagta actgctgaga ccgaattctc gccctatagt
      901 gagtcgtatt acctggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      961 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1021 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     1081 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     1141 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     1201 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1261 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     1321 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1381 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1501 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1621 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1681 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1741 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1801 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1921 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2041 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2101 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2221 ggctctcgag gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2341 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3241 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3301 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3361 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3421 agttagctca ctcat
//
