LOCUS       sequence-331866-        7669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mGreenLantern to check for monomericity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7669)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 7669)
  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: mGreenLantern pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(109..564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(246..267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     458..477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1372..1396
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1403..1405,1406..1408,1409..2122)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(1448..1469)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2131..2155)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2217..2236)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2218..2236)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2384..2571
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2439..2463)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2583..2600)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2583..2599)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2592..2614)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3035..4381
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4492..5295)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5296..5511)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5536..5554)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5622..5726
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5727..5795,5796..6587)
                     /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(5945..5964)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6758..7346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7247..7266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7500..7517
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7634..7655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagcttacc gcatcaggaa attgtaagcg
      121 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
      181 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      241 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
      301 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      361 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      421 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      481 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      541 ttaatgcgcc gctacagggc gcgtccattc gccaagcttc ctgaaacgga gaaacataaa
      601 caggcattgc tgggatcacc catacatcac tctgttttgc ctgacctttt ccggtaattt
      661 gaaaacaaac ccggtctcga agcggagatc cggcgataat taccgcagaa ataaacccat
      721 acacgagacg tagaaccagc cgcacatggc cggagaaact cctgcgagaa tttcgtaaac
      781 tcgcgcgcat tgcatctgta tttcctaatg cggcacttcc aggcctcgat cgagaccgtt
      841 tatccattgc ttttttgttg tctttttccc tcgttcacag aaagtctgaa gaagctatag
      901 tagaactatg agcttttttt gtttctgttt tccttttttt tttttttacc tctgtggaaa
      961 ttgttactct cacactcttt agttcgtttg tttgttttgt ttattccaat tatgaccggt
     1021 gacgaaacgt ggtcgatggt gggtaccgct tatgctcccc tccattagtt tcgattatat
     1081 aaaaaggcca aatattgtat tattttcaaa tgtcctatca ttatcgtcta acatctaatt
     1141 tctcttaaat tttttctctt tctttcctat aacaccaata gtgaaaatct ttttttcttc
     1201 tatatctaca aaaacttttt ttttctatca acctcgttga taaatttttt ctttaacaat
     1261 cgttaataat taattaattg gaaaataacc attttttctc tcttttatac acacattcaa
     1321 aagaaagaaa aaaaatatac cccagcctcg actagtgaat tcgcggccat cacaagtttg
     1381 tacaaaaaag caggctgcta gcatggtgag caagggcgag gagctgttca ccggggtggt
     1441 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga
     1501 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1561 gctgcccgtg ccctggccca ccctcgtgac caccttaggc tacggcgtgg cctgcttcgc
     1621 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1681 cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt
     1741 gaagttcgag ggcgacaccc tggtgaaccg catcgtgctg aagggcatcg acttcaagga
     1801 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca aggtctatat
     1861 cacggccgac aagcagaaga acggcatcaa ggctaacttc aagacccgcc acaacgttga
     1921 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1981 cgtgctgctg cccgacaacc actacctgag ccatcagtcc aaactgagca aagaccccaa
     2041 cgagaagcgc gatcacatgg tcctgaagga gagggtgacc gccgccggga ttacacatga
     2101 catggacgag ctgtacaagt aagcggccgc acccagcttt cttgtacaaa gtggtgtagt
     2161 taattcatga tggccgctgc aggtcgacct cgaggggggg cccggtaccc aattcgccct
     2221 atagtgagtc gtattacgcg cggatccagc tttggacttc ttcgccagag gtttggtcaa
     2281 gtctccaatc aaggttgtcg gcttgtctac cttgccagaa atttacgaaa agatggaaaa
     2341 gggtcaaatc gttggtagat acgttgttga cacttctaaa taagcgaatt tcttatgatt
     2401 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2461 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2521 gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgccaat
     2581 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2641 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2701 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2761 cttacgcatc tgtgcggtat ttcacaccgc ataatcggat cgtacttgtt acccatcatt
     2821 gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat ttgaacctgt
     2881 ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg ttcctggaga
     2941 aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc attttctgcg
     3001 tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc ttcattttgt
     3061 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     3121 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     3181 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     3241 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     3301 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     3361 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     3421 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     3481 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     3541 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     3601 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     3661 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     3721 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     3781 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     3841 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     3901 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     3961 gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     4021 gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa
     4081 atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt
     4141 gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta
     4201 cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc
     4261 cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct
     4321 gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat
     4381 cgatccgatg ataagctgtc aaacatgaga attgggtaat aactgatata attaaattga
     4441 agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg
     4501 ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac
     4561 cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt
     4621 agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa
     4681 acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct
     4741 ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt
     4801 agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc
     4861 tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc
     4921 caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga
     4981 gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa
     5041 attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt
     5101 caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc
     5161 cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg
     5221 catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata
     5281 tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt
     5341 taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct
     5401 aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt
     5461 tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctaatt
     5521 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5581 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5641 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5701 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5761 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5821 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5881 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5941 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6001 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6061 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6121 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6181 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6241 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6301 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6361 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6421 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6481 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6541 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6601 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6661 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6721 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6781 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6841 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6901 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6961 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7021 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7081 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7141 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7201 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7261 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7321 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7381 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7441 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7501 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7561 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7621 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggc
//
