LOCUS       sequence-320901-        6120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acceptor for golden gate assembly with BsaI overhangs GCTT-CCAT. 
            Generates plasmids for T7-driven expression in E. coli. Contains 
            mRFP1 reporter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6120)
  AUTHORS   Dudley QM, Cai YM, Kallam K, Debreyne H, Carrasco Lopez JA, Patron 
            NJ
  TITLE     Biofoundry-assisted expression and characterization of plant 
            proteins.
  JOURNAL   Synth Biol (Oxf). 2021 Sep 11;6(1):ysab029. doi: 
            10.1093/synbio/ysab029. eCollection 2021.
  PUBMED    34693026
REFERENCE   2  (bases 1 to 6120)
  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: "Synth Biol 
            (Oxf)."; date: "2021-09-11"; volume: "6(1):ysab029. doi"; pages: " 
            10.1093/synbio/ysab029. eCollection 2021"
COMMENT     Sequence Label: pEPMY1CB0001
FEATURES             Location/Qualifiers
     source          1..6120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     371..393
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     455..474
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     516..535
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        516..534
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    535..559
                     /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        575..684
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     protein_bind    821..842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(857..862,863..880,881..887)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    895..911
                     /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)."
     RBS             929..940
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             947..1624
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1669..1740
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1756..1783
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1906..1925)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1971..2026
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2025..2044)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2114..2161
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2174..2879
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2262..2634
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2291..2879)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2888..2921)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3089..3676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3168..3187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3689..4480,4481..4549))
                     /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     4312..4331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     4792..5619
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4792..4916
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4954..5559)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5751..6054
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
       61 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
      121 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
      181 cacaggtgag cgggcgggac ggcccttctc cttcgggctg taattagcgc ttggtttaat
      241 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
      301 tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
      361 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
      421 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
      481 catcattttg gcaaagaatt ggatcggacc gaaattaata cgactcacta taggggaatt
      541 gtgagcggat aacaattccc cggagttaat ccgggacctt taattcaacc caacacaata
      601 tattatagtt aaataagaat tattatcaaa tcatttgtat attaattaaa atactatact
      661 gtaaattaca ttttatttac aatcaaagga gatataccga attcccattg agaccgatat
      721 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      781 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      841 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      901 gcggataaca atttcacaca tactagagaa agaggagaaa tactagatgg cttcctccga
      961 ggacgttatc aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca
     1021 cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa
     1081 actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt
     1141 ccagtacggt tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact
     1201 gtccttcccg gaaggtttca aatgggaacg tgttatgaac ttcgaggacg gtggtgttgt
     1261 taccgttacc caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg
     1321 tggtaccaac ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc
     1381 ttccaccgaa cgtatgtacc cggaggacgg tgctctgaaa ggtgaaatca aaatgcgtct
     1441 gaaactgaaa gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa
     1501 accggttcag ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa
     1561 cgaggactac accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc
     1621 ttaataacgc tgatagtgct agtgtagatc gctactagag ccaggcatca aataaaacga
     1681 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1741 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag gtctcagctt
     1801 tctagggatc caccagtttg tgattaacct caggtgcagg ctgcctatca gaaggtggtg
     1861 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atcgatcttt ttccctctgc
     1921 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     1981 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2041 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     2101 tgcccatatg taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2161 gctgaaagca tgcggaggaa attctccttg aagtttccct ggtgttcaaa gtaaaggagt
     2221 ttgcaccaga cgcacctctg ttcactggtc cggcgtatta aaacacgata cattgttatt
     2281 agtacattta ttaagcgcta gattctgtgc gttgttgatt tacagacaat tgttgtacgt
     2341 attttaataa ttcattaaat ttataatctt tagggtggta tgttagagcg aaaatcaaat
     2401 gattttcagc gtctttatat ctgaatttaa atattaaatc ctcaatagat ttgtaaaata
     2461 ggtttcgatt agtttcaaac aagggttgtt tttccgaacc gatggctgga ctatctaatg
     2521 gattttcgct caacgccaca aaacttgcca aatcttgtag cagcaatcta gctttgtcga
     2581 tattcgtttg tgttttgttt tgtaataaag gttcgacgtc gttcaaaata ttatgcgctt
     2641 ttgtatttct ttcatcactg tcgttagtgt acaattgact cgacgtaaac acgttaaata
     2701 gagcttggac atatttaaca tcgggcgtgt tagctttatt aggccgatta tcgtcgtcgt
     2761 cccaaccctc gtcgttagaa gttgcttccg aagatgattt tgccatagcc acacgacgcc
     2821 tattaattgt gtcggctaac acgtccgcga tcaaatttgt agttgagctt tttggaattg
     2881 cgatcgcata acttcgtata gcatacatta tacgaagtta taagctcgga acgctgcgct
     2941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgccccctg acgagcatca
     3121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3661 aaaaaaagga tctcaagaag atcctttgtt accaatgctt aatcagtgag gcacctatct
     3721 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3781 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     3841 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3901 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3961 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4021 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4081 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4141 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4201 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4261 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4321 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4381 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4441 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4501 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4561 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gtccgcgcgt ttcctgcatc
     4621 ttttaatcaa atcccaagat gtgtataaac gcgccggtat gtacaggaag aggtttatac
     4681 taaactgtta cattgcaaac gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca
     4741 aggctctgac gcatttctac aaccacgact ccaagtgtgt gggtgaagtc atgcatcttt
     4801 taatcaaatc ccaagatgtg tataaaccac caaactgcca aaaaatgaaa actgtcgaca
     4861 agctctgtcc gtttgctggc aactgcaagg gactcaatcc tatttgtaat tattgaataa
     4921 taaaacaatt ataaatgtca aatttgtttt ttattaacga tacaaaccaa acgcaacaag
     4981 aacatttgta gtattatcta taattgaaaa cgcgtagtta taatcgctga ggtaatattt
     5041 aaaatcattt tcaaatgatt cacagttaat ttgcgacaat ataattttat tttcacataa
     5101 actagacgcc ttgtcgtcct cttcttcgta ttccttctct ttttcatttt tctcttcata
     5161 aaaattaaca tagttattat cgtatccata tatgtatcta tcgtatagag taaatttttt
     5221 gttgtcataa atatatatgt cttttttaat ggggtgtata gtaccgctgc gcatagtttt
     5281 tctgtaattt acaacagtgc tattttctgg tagttcttcg gagtgtgttg ctttaattat
     5341 taaatttata taatcaatga atttgggatc gtcggttttg tacaatatgt tgccggcata
     5401 gtacgcagct tcttctagtt caattacacc attttttagc agcaccggat taacataact
     5461 ttccaaaatg ttgtacgaac cgttaaacaa aaacagttca cctccctttt ctatactatt
     5521 gtctgcgagc agttgtttgt tgttaaaaat aacagccatt gtaatgagac gcacaaacta
     5581 atatcacaaa ctggaaatgt ctatcaatat atagttgctg atggccggcc gtaatgagac
     5641 gcacaaacta atatcacaaa ctggaaatgt ctatcaatat atagttgctc tagttattaa
     5701 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     5761 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5821 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5881 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5941 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6001 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgcatggt
     6061 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
//
