LOCUS       sequence-269096-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Goldengate/Greengate module marker for transgenic Arabidopsis seed 
            coat fluorescence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5797)
  TITLE     Theres lab:  Alternative Greengate/Goldengate modules
REFERENCE   2  (bases 1 to 5797)
  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"
COMMENT     Sequence Label: pGGF_RT_FAST_Tq2CFP
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     promoter        2212..2230
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2212..2229
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(4393..4395,4396..4398,4399..5112)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4438..4459)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4699..4718)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5046..5067
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5114..5362
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     primer_bind     complement(5383..5402)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5384..5402)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5412..5434)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5628..5647)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5747..5769
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtggctct cgaggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacgagc tatttaggtg
     2221 acactataga agtgaagctt ggtctcaact acttcaagtg tatgtaggta tagtaacatg
     2281 aacaagaaca gactcaagta caagatcgca tacgaaaatg gaaatggcaa tgtcacttcc
     2341 acataatcaa acacgaatcc tcatatcaac aaggcctgag attctaacta gctcataaca
     2401 acttagccaa tagttacttg agactaccaa atgtatgtag aactaaagac taagggacag
     2461 agagttcgtc taaacaggtg aatctagtcg ttgttatcta ataaacaatt cagccccaaa
     2521 tgcagaacac acatagagct ctctattgat tcaaattacg atctgatact gataacgtct
     2581 agatttttag ggttaaagca atcaatcacc tgacgattca aggtggttgg atcatgacga
     2641 ttccagaaaa catcaagcaa gctctcaaag ctacactctt tgggatcata ctgaactcta
     2701 acaacctcgt tatgtcccgt agtgccagta cagacatcct cgtaactcgg attgtgcacg
     2761 atgccatggc tatacccaac ctcggtcttg gtcacaccag gaactctctg gtaagctagc
     2821 tccactcccc agaaacaacc ggcgccaaat tgcgcgaatt gctgacctga agacggaaca
     2881 tcatcgtcgg gtccttgggc gattgcggcg gaagatgggt cagcttgggc ttgaggacaa
     2941 gacccgaatc cgagtctgtt gaaaaggttg ttcattgggg atttgtatac ggagattggt
     3001 cgtcgagagg tttgagggaa aggacaaatg ggtttggctc tggagaaaga gagtgcggct
     3061 ttagagagag aattgagagg tttagagaga gatgcggcgg cgatgagcgg aggagagacg
     3121 acgaggacct gcattatcaa agcagtgacg tggtgaaatt tggaactttt aagaggcaga
     3181 tagatttatt atttgtatcc attttcttca ttgttctaga atgtcgcgga acaaatttta
     3241 aaactaaatc ctaaattttt ctaattttgt tgccaatagt ggatatgtgg gccgtataga
     3301 aggaatctat tgaaggccca aacccatact gacgagccca aaggttcgtt ttgcgtttta
     3361 tgtttcggtt cgatgccaac gccacattct gagctaggca aaaaacaaac gtgtctttga
     3421 atagactcct ctcgttaaca catgcagcgg ctgcatggtg acgccattaa cacgtggcct
     3481 acaattgcat gatgtctcca ttgacacgtg acttctcgtc tcctttctta atatatctaa
     3541 caaacactcc tacctcttcc aaaatatata cacatctttt tgatcaatct ctcattcaaa
     3601 atctcattct ctctagtaaa caagaacaaa aaaatggcgg atacagctag aggaacccat
     3661 cacgatatca tcggcaggga ccagtacccg atgatgggcc gagatcggga ccagtaccag
     3721 atgtccggac gaggatctga ctactccaag tctaggcaga ttgctaaagc tgcaactgct
     3781 gtcacagctg gtggttccct ccttgttctc tccagcctta cccttgttgg aactgtcata
     3841 gctttgactg ttgcaacacc tctgctcgtt atcttcagcc caatccttgt cccggctctc
     3901 atcacagttg cactcctcat caccggtttt ctttcctctg gagggtttgg cattgccgct
     3961 ataaccgttt tctcttggat ttacaagtaa gcacacattt atcatcttac ttcataattt
     4021 tgtgcaatat gtgcatgcat gtgttgagcc agtagctttg gatcaatttt tttggtcgaa
     4081 taacaaatgt aacaataaga aattgcaaat tctagggaac atttggttaa ctaaatacga
     4141 aatttgacct agctagcttg aatgtgtctg tgtatatcat ctatataggt aaaatgcttg
     4201 gtatgatacc tattgattgt gaataggtac gcaacgggag agcacccaca gggatcagac
     4261 aagttggaca gtgcaaggat gaagttggga agcaaagctc aggatctgaa agacagagct
     4321 cagtactacg gacagcaaca tactggtggg gaacatgacc gtgaccgtac tcgtggtggc
     4381 cagcacacta ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     4441 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     4501 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     4561 ccctggccca ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc
     4621 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     4681 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     4741 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     4801 atcctggggc acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac
     4861 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     4921 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4981 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     5041 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     5101 ctgtacaagt gaatatgaag atgaagatga aatatttggt gtgtcaaata aaaagcttgt
     5161 gtgcttaagt ttgtgttttt ttcttggctt gttgtgttat gaatttgtgg ctttttctaa
     5221 tattaaatga atgtaagatc tcattataat gaataaacaa atgtttctat aatccattgt
     5281 gaatgttttg ttggatctct tctgcagcat ataactactg tatgtgctat ggtatggact
     5341 atggaatatg attaaagata aggtattgag accgaattct cgccctatag tgagtcgtat
     5401 tacctggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5461 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5521 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     5581 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     5641 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5701 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5761 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
