LOCUS       sequence-314659-        6245 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 6245)
  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 6245)
  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..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     63..82
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        63..81
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    82..106
                     /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        122..231
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     protein_bind    368..389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(404..409,410..427,428..434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    442..458
                     /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             476..487
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             494..1171
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1216..1287
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1303..1330
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1453..1472)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1518..1573
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1572..1591)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      1661..1708
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     1721..2426
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           1809..2181
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(1838..2426)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2435..2468)
                     /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(2636..3223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2715..2734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3236..4027,4028..4096))
                     /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     3859..3878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     4339..5166
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4339..4463
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4501..5106)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5298..5601
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5607..5884
                     /label=chicken beta-actin promoter
     primer_bind     6163..6185
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tcggaccgaa
       61 attaatacga ctcactatag gggaattgtg agcggataac aattccccgg agttaatccg
      121 ggacctttaa ttcaacccaa cacaatatat tatagttaaa taagaattat tatcaaatca
      181 tttgtatatt aattaaaata ctatactgta aattacattt tatttacaat caaaggagat
      241 ataccgaatt cccattgaga ccgatatcaa tacgcaaacc gcctctcccc gcgcgttggc
      301 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      361 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
      421 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacatac tagagaaaga
      481 ggagaaatac tagatggctt cctccgagga cgttatcaaa gagttcatgc gtttcaaagt
      541 tcgtatggaa ggttccgtta acggtcacga gttcgaaatc gaaggtgaag gtgaaggtcg
      601 tccgtacgaa ggtacccaga ccgctaaact gaaagttacc aaaggtggtc cgctgccgtt
      661 cgcttgggac atcctgtccc cgcagttcca gtacggttcc aaagcttacg ttaaacaccc
      721 ggctgacatc ccggactacc tgaaactgtc cttcccggaa ggtttcaaat gggaacgtgt
      781 tatgaacttc gaggacggtg gtgttgttac cgttacccag gactcctccc tgcaagacgg
      841 tgagttcatc tacaaagtta aactgcgtgg taccaacttc ccgtccgacg gtccggttat
      901 gcagaaaaaa accatgggtt gggaagcttc caccgaacgt atgtacccgg aggacggtgc
      961 tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac ggtggtcact acgacgctga
     1021 agttaaaacc acctacatgg ctaaaaaacc ggttcagctg ccgggtgctt acaaaaccga
     1081 catcaaactg gacatcacct cccacaacga ggactacacc atcgttgaac agtacgaacg
     1141 tgctgaaggt cgtcactcca ccggtgctta ataacgctga tagtgctagt gtagatcgct
     1201 actagagcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1261 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     1321 cctttctgcg tttataggtc tcagctttct agggatccac cagtttgtga ttaacctcag
     1381 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     1441 cactgagatc gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     1501 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     1561 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     1621 gtatttggtt tagagtttgg caacatatgc ccatatgtaa ctagcataac cccttggggc
     1681 ctctaaacgg gtcttgaggg gttttttgct gaaagcatgc ggaggaaatt ctccttgaag
     1741 tttccctggt gttcaaagta aaggagtttg caccagacgc acctctgttc actggtccgg
     1801 cgtattaaaa cacgatacat tgttattagt acatttatta agcgctagat tctgtgcgtt
     1861 gttgatttac agacaattgt tgtacgtatt ttaataattc attaaattta taatctttag
     1921 ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc tttatatctg aatttaaata
     1981 ttaaatcctc aatagatttg taaaataggt ttcgattagt ttcaaacaag ggttgttttt
     2041 ccgaaccgat ggctggacta tctaatggat tttcgctcaa cgccacaaaa cttgccaaat
     2101 cttgtagcag caatctagct ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt
     2161 cgacgtcgtt caaaatatta tgcgcttttg tatttctttc atcactgtcg ttagtgtaca
     2221 attgactcga cgtaaacacg ttaaatagag cttggacata tttaacatcg ggcgtgttag
     2281 ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc gttagaagtt gcttccgaag
     2341 atgattttgc catagccaca cgacgcctat taattgtgtc ggctaacacg tccgcgatca
     2401 aatttgtagt tgagcttttt ggaattgcga tcgcataact tcgtatagca tacattatac
     2461 gaagttataa gctcggaacg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2521 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2581 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2641 ataggctccg ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2701 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2761 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2821 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2881 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2941 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3001 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3061 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3121 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3181 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgttacc
     3241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3361 ctgcaatgat accgcgagat ccacgctcac cggctccaga tttatcagca ataaaccagc
     3421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4141 gtctcatgtc cgcgcgtttc ctgcatcttt taatcaaatc ccaagatgtg tataaacgcg
     4201 ccggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg gtttcgtgtg
     4261 ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac cacgactcca
     4321 agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat aaaccaccaa
     4381 actgccaaaa aatgaaaact gtcgacaagc tctgtccgtt tgctggcaac tgcaagggac
     4441 tcaatcctat ttgtaattat tgaataataa aacaattata aatgtcaaat ttgtttttta
     4501 ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa ttgaaaacgc
     4561 gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac agttaatttg
     4621 cgacaatata attttatttt cacataaact agacgccttg tcgtcctctt cttcgtattc
     4681 cttctctttt tcatttttct cttcataaaa attaacatag ttattatcgt atccatatat
     4741 gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt ttttaatggg
     4801 gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat tttctggtag
     4861 ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt tgggatcgtc
     4921 ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa ttacaccatt
     4981 ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt taaacaaaaa
     5041 cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt taaaaataac
     5101 agccattgta atgagacgca caaactaata tcacaaactg gaaatgtcta tcaatatata
     5161 gttgctgatg gccggccgta atgagacgca caaactaata tcacaaactg gaaatgtcta
     5221 tcaatatata gttgctctag ttattaatag taatcaatta cggggtcatt agttcatagc
     5281 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     5341 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     5401 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     5461 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     5521 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     5581 ttagtcatcg ctattaccat gcatggtcga ggtgagcccc acgttctgct tcactctccc
     5641 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     5701 agcgatgggg gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
     5761 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     5821 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     5881 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     5941 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     6001 tccttcgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     6061 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggctgtc
     6121 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     6181 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     6241 tcctg
//
