LOCUS       sequence-332096-        7868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cyterm-mGreenLantern to check for monomericity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7868)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 7868)
  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: Cyterm-mGreenLantern pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..7868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     414..435
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      477..486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(686..710)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      1518..1973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1605..1624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1815..1836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2003..2019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2003..2019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2016..2038)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2027..2043
                     /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        complement(join(2051..2057,2058..2075,2076..2081))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2096..2117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2234..2251)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2405..2993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2485..2504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3164..3955,3956..4024))
                     /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     3787..3806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4025..4129)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4197..4215
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4240..4455
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4456..5259
                     /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"
     rep_origin      5370..6716
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     7137..7159
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7151..7168
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7152..7168
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(7180..7367)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     7288..7312
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     7515..7534
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7515..7533
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7596..7620
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(7629..7868,1..474,475..477,478..480))
                     /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"
ORIGIN
        1 gttcttctgc ttgtcggccg tgatatagac cttgtggctg ttgaagttgt actccagctt
       61 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagcacga tgcggttcac
      121 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg taggtaccgt cgtccttgaa
      181 agagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
      241 cttcatgtgg tcggggtagc gggcgaagca ggccacgccg tagcctaagg tggtcacgag
      301 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
      361 gttggtggca tcgccctcgc cctcgccgcg gacgctgaac ttgtggccgt ttacgtcgcc
      421 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
      481 ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc gaagcttccc
      541 tcccccatag ctctgtttcc agagtgaaag gagaagcaaa caggagagac agagccccag
      601 caccaccaca ggatccatgg gagccttctc ttcctgacaa ctggaagctg caaggggtac
      661 cagatctgag tccggtagcg ctagcagcct gcttttttgt acaaacttgt gatggccgcg
      721 aattcactag tcgaggctgg ggtatatttt ttttctttct tttgaatgtg tgtataaaag
      781 agagaaaaaa tggttatttt ccaattaatt aattattaac gattgttaaa gaaaaaattt
      841 atcaacgagg ttgatagaaa aaaaaagttt ttgtagatat agaagaaaaa aagattttca
      901 ctattggtgt tataggaaag aaagagaaaa aatttaagag aaattagatg ttagacgata
      961 atgataggac atttgaaaat aatacaatat ttggcctttt tatataatcg aaactaatgg
     1021 aggggagcat aagcggtacc caccatcgac cacgtttcgt caccggtcat aattggaata
     1081 aacaaaacaa acaaacgaac taaagagtgt gagagtaaca atttccacag aggtaaaaaa
     1141 aaaaaaaagg aaaacagaaa caaaaaaagc tcatagttct actatagctt cttcagactt
     1201 tctgtgaacg agggaaaaag acaacaaaaa agcaatggat aaacggtctc gatcgaggcc
     1261 tggaagtgcc gcattaggaa atacagatgc aatgcgcgcg agtttacgaa attctcgcag
     1321 gagtttctcc ggccatgtgc ggctggttct acgtctcgtg tatgggttta tttctgcggt
     1381 aattatcgcc ggatctccgc ttcgagaccg ggtttgtttt caaattaccg gaaaaggtca
     1441 ggcaaaacag agtgatgtat gggtgatccc agcaatgcct gtttatgttt ctccgtttca
     1501 ggaagcttgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1561 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1621 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1681 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1741 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1801 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1861 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     1921 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     1981 cggtaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2041 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2101 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2161 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2221 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2281 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2341 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2401 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2461 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2521 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2581 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2641 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2701 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2761 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2821 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2881 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2941 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3001 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3061 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3121 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3181 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3241 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3301 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3361 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3421 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3481 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3541 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3601 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3661 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3721 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3781 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3841 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3901 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3961 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4021 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4081 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4141 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4201 ggcgtatcac gaggcccttt cgtcttcaag aattagcttt tcaattcatc attttttttt
     4261 tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag
     4321 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     4381 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacatgcagg
     4441 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     4501 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     4561 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     4621 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     4681 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct
     4741 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     4801 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     4861 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     4921 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     4981 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     5041 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa
     5101 cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga
     5161 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     5221 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     5281 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccaat
     5341 tctcatgttt gacagcttat catcggatcg atccaatatc aaaggaaatg atagcattga
     5401 aggatgagac taatccaatt gaggagtggc agcatataga acagctaaag ggtagtgctg
     5461 aaggaagcat acgatacccc gcatggaatg ggataatatc acaggaggta ctagactacc
     5521 tttcatccta cataaataga cgcatataag tacgcattta agcataaaca cgcactatgc
     5581 cgttcttctc atgtatatat atatacaggc aacacgcaga tataggtgcg acgtgaacag
     5641 tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt
     5701 tgaagttcct attccgaagt tcctattctc tagctagaaa gtataggaac ttcagagcgc
     5761 ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac gcaccggact
     5821 gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca aaagtatctc
     5881 tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct ttcgctcctt
     5941 gaacttgcat ctaaactcga cctctacatt ttttatgttt atctctagta ttactcttta
     6001 gacaaaaaaa ttgtagtaag aactattcat agagtgaatc gaaaacaata cgaaaatgta
     6061 aacatttcct atacgtagta tatagagaca aaatagaaga aaccgttcat aattttctga
     6121 ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa agtatgcgca atccacatcg
     6181 gtatagaata taatcgggga tgcctttatc ttgaaaaaat gcacccgcag cttcgctagt
     6241 aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa tagacaccaa
     6301 agtagccttc ttctaacctt aacggaccta cagtgcaaaa agttatcaag agactgcatt
     6361 atagagcgca caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa aatagcgctc
     6421 tcgggatgca tttttgtaga acaaaaaaga agtatagatt ctttgttggt aaaatagcgc
     6481 tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag
     6541 cgctctcgcg ttgcattttt gttttacaaa aatgaagcac agattcttcg ttggtaaaat
     6601 agcgctttcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa
     6661 attagcgctc tcgcgttgca tttttgttct acaaaatgaa gcacagatgc ttcgttaaca
     6721 aagatatgct attgaagtgc aagatggaaa cgcagaaaat gaaccgggga tgcgacgtgc
     6781 aagattacct atgcaataga tgcaatagtt tctccaggaa ccgaaataca tacattgtct
     6841 tccgtaaagc gctagactat atattattat acaggttcaa atatactatc tgtttcaggg
     6901 aaaactccca ggttcggatg ttcaaaattc aatgatgggt aacaagtacg atccgattat
     6961 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     7021 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     7081 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     7141 gtcacgacgt tgtaaaacga cggccagtga attggcatgc cggtagaggt gtggtcaata
     7201 agagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag
     7261 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt
     7321 tttataactt atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg
     7381 tcaacaacgt atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag
     7441 gtagacaagc cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaagtccaaa
     7501 gctggatccg cgcgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg
     7561 aggtcgacct gcagcggcca tcatgaatta actacaccac tttgtacaag aaagctgggt
     7621 gcggccgctt acttgtacag ctcgtccatg tcatgtgtaa tcccggcggc ggtcaccctc
     7681 tccttcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagttt ggactgatgg
     7741 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     7801 tagtggtcgg cgagctgcac gccgccgtcc tcaacgttgt ggcgggtctt gaagttagcc
     7861 ttgatgcc
//
