LOCUS       sequence-490747-        7321 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  CaMV 35S-SynPro-13 engineered promoter driving the expression of 
            Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7321)
  AUTHORS   Khan MA, Herring G, Zhu JY, Oliva M, Fourie E, Johnston B, Zhang Z, 
            Potter J, Pineda L, Pflueger J, Swain T, Pflueger C, Lloyd JPB, 
            Secco D, Small I, Kidd BN, Lister R
  TITLE     CRISPRi-based circuits to control gene expression in plants.
  JOURNAL   Nat Biotechnol. 2025 Mar;43(3):416-430. doi: 
            10.1038/s41587-024-02236-w. Epub 2024 May 20.
  PUBMED    38769424
REFERENCE   2  (bases 1 to 7321)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41587-024-02236-w"; journalName: "Nat Biotechnol"; date: 
            "2025-03"; volume: "43"; issue: "3"; pages: "416-430"
COMMENT     Sequence Label: LLP978
FEATURES             Location/Qualifiers
     source          1..7321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1610..1798
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     1798..1816
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(1893..2072)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    2086..2177
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     polyA_signal    2191..2239
                     /note="synthetic polyadenylation signal"
     intron          3471..3659
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     primer_bind     complement(4100..4119)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4101..4119)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4126..4143)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4126..4142)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4135..4157)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4284..4739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4371..4390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4581..4602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4765..4869
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4870..4938,4939..5730)
                     /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(5088..5107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5901..6489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6390..6409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6643..6660
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6777..6798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6813..6818,6819..6836,6837..6843)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6851..6867
                     /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     6856..6878
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6875..6891
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6875..6891
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6912..6930
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    6949..7040
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     polyA_signal    7054..7102
                     /note="synthetic polyadenylation signal"
     terminator      complement(join(7103..7321,1..34))
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 ttaagaaact ttattgccaa atgtttgaac gatcctcgag ttacacggcg atctttccgc
       61 ccttcttggc ctttatgagg atctctctga tttttcttgc gtcgagtttt ccggtaagac
      121 ctttcggtac ttcgtccaca aacacaactc ctccgcgcaa ctttttcgcg gttgttactt
      181 gactggcgac gtaatccacg atctcttttt ccgtcatcgt ctttccgtgc tccaaaacaa
      241 caacggcggc gggaagttca ccggcgtcat cgtcgggaag acctgcgaca cctgcgtcga
      301 agatgttggg gtgttggagc aagatggatt ccaattcagc gggagccacc tgatagcctt
      361 tgtacttaat cagagacttc aggcggtcaa cgatgaagaa gtgttcgtct tcgtcccagt
      421 aagctatgtc tccagaatgt agccatccat ccttgtcaat caaggcgttg gtcgcttccg
      481 gattgtttac ataaccggac ataatcatag gacctctcac acacagttcg cctctttgat
      541 taacgcccag cgttttcccg gtatccagat ccacaacctt cgcttcaaaa aatggaacaa
      601 ctttaccgac cgcgcccggt ttatcatccc cctcgggtgt aatcagaata gctgatgtag
      661 tctcagtgag cccatatcct tgcctgatac ctggcagatg gaacctcttg gcaaccgctt
      721 ccccgacttc cttagagagg ggagcgccac cagaagcaat ttcgtgtaaa ttagataaat
      781 cgtatttgtc aatcagagtg cttttggcga agaaggagaa tagggttggc accagcagcg
      841 cactttgaat cttgtaatcc tgaaggctcc tcagaaacag ctcttcttca aatctataca
      901 ttaagacgac tcgaaatcca catatcaaat atccgagtgt agtaaacatt ccaaaaccgt
      961 gatggaatgg aacaacactt aaaatcgcag tatccggaat gatttgattg ccaaaaatag
     1021 gatctctggc atgcgagaat ctcacgcagg cagttctatg aggcagagcg acacctttag
     1081 gcagaccagt agatccagag gagttcatga tcagtgcaat tgtcttgtcc ctatcgaagg
     1141 actctggcac aaaatcgtat tcattaaaac cgggaggtag atgagatgtg acgaacgtgt
     1201 acatcgactg aaatccctgg taatccgttt tagaatccat gataataatt ttttggatga
     1261 ttgggagctt tttttgcacg ttcaaaattt tttgcaaccc ctttttggaa acgaacacca
     1321 cggtaggctg cgaaatgccc atactgttga gcaattcacg ttcattataa atgtcgttcg
     1381 cgggcgcaac tgcaactccg ataaataacg cgcccaacac cggcataaag aattgaagag
     1441 agttttcact gcatacgacg attctgtgat ttgtattcag cccatatcgt ttcatagctt
     1501 ctgccaaccg aacggacatt tcgaagtact cagcgtaagt gatgtccacc tcgatatgtg
     1561 catctgtaaa agcaattgtt ccaggaacca gggcgtatct cttcatagcc tgcacatcaa
     1621 caaattttgg tcatatatta gaaaagttat aaattaaaat atacacactt ataaactaca
     1681 gaaaagcaat tgctatatac tacattcttt tattttgaaa aaaatatttg aaatattata
     1741 ttactactaa ttaatgataa ttattatata tatatcaaag gtagaagcag aaacttacct
     1801 tatgcagttg ctctccagcg gttccatctt ccagcggata gaatggcgcc gggcctttct
     1861 ttatgttttt ggcgtcttcc atccatggac tctaattgga taccgagggg aatttatgga
     1921 acgtcagtgg agcatttttg acaagaaata tttgctagct gatagtgacc ttaggcgact
     1981 tttgaacgcg caataatggt ttctgacgta tgtgcttagc tcattaaact ccagaaaccc
     2041 gcggctcagt ggctccttca acgttgcggt tctctagaga gcgctagaga aatgttctgg
     2101 cacctgcact tgcactgggg acagcctatt ttgctagttt gttttgtttc gttttgtttt
     2161 gatggagagc gtatgttagt actatcgatt cacacaaaaa accaacacac agatgtaatg
     2221 aaaataaaga tattttatta ctagtaggtc actggatttt ggttttagga attagaaatt
     2281 ttattgatag aagtatttta caaatacaaa tacatactaa gggtttctta tatgctcaac
     2341 acatgagcga aaccctataa gaaccctaat tcccttatct gggaactact cacacattat
     2401 tctggagaaa aatagagaga gatagatttg tagagagaga ctggtgattt ttgcggactc
     2461 tagcatggcc gggtacctta agaaccaaca caacctggcg tttccactgt gcatacctca
     2521 gactgggagc ttgttgacgt tacttcagaa gcatcagatc gagaaccctg ctcgtttttg
     2581 agcactcttt caacaaaaga cttaatatac tttcccatct catcaggagc atcttcttgt
     2641 gagaaatgca atcctttcac cttaacgaac tcggtgtttg gaaatttctt tgcaccttca
     2701 actatagcat tactgaaaaa tcctggatct gactcgatga acatcttagg caaatcatct
     2761 gatgccctaa ggtaagcgtt ataatttctc acgatctgca caacatcagg tttacctccc
     2821 ttaactaatg gtatttccct aggccaactg agtgttggcc ttctaacctc tcctttttcc
     2881 ttgaaaggtt cgagatatgc agcaaactct tctggttcca attttctcat aatctttgaa
     2941 ggaagcatag tttccacgaa aaagttattt tcgagaacca tcttttctcc ctcttcagac
     3001 ttgatcaaag caatatcctc ttcaatatct ggccactcat cccaagattc tatcacatcc
     3061 acaacactct ctgcatgaac tatagctttg atcttatctt ggtgttcgta tgagtaatgg
     3121 aatgccaagc aagcacccca atcgtgtcca acgaaaataa tcttcttagg taagttgaga
     3181 agttcgaacc aagctgtgag gtacttataa tgatccaaaa gtctgtaaga accatttcca
     3241 gatttaccag actttcccat tccgataaga tctgggataa tacacctagc cactggttca
     3301 atgtgaggca caacatgtct ccacaaatat gaagatgcag cgtttccgtg aagaaagata
     3361 actgcattct cagcgtgctt ttctgaatcg tagtaattaa tgaaagaatc aagcacgttc
     3421 atctgtttac atcttgccca ccattgtgga ccagttatca ttctttttct ctgcacatca
     3481 acaaattttg gtcatatatt agaaaagtta taaattaaaa tatacacact tataaactac
     3541 agaaaagcaa ttgctatata ctacattctt ttattttgaa aaaaatattt gaaatattat
     3601 attactacta attaatgata attattatat atatatcaaa ggtagaagca gaaacttacc
     3661 tgctcaggat catacacctt tgaagtcatc tgcagttatg tcgacggatc cgtccccatt
     3721 cgtctgccta tggatgctta tataccacca caaccggctg agctgccgtg ggattgtgcg
     3781 tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga cgtggttgga
     3841 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
     3901 gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt gtaggtgcca
     3961 ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg gaatccgagg
     4021 aggtttcccg atattaccct ttgttgaaaa gtctcaaacg cgttcggaat tcggcatgat
     4081 caagcttgta cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
     4141 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4201 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4261 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4441 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4621 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4681 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     4741 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4801 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4861 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4921 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4981 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5041 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5101 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5161 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5221 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5281 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5341 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5401 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5461 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5521 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5581 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5641 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5701 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5761 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5821 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5881 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5941 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6001 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6061 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6121 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6181 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6241 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6301 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6361 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6421 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6481 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6541 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6601 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6661 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6721 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6781 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6841 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6901 gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcag agaaatgttc
     6961 tggcacctgc acttgcactg gggacagcct attttgctag tttgttttgt ttcgttttgt
     7021 tttgatggag agcgtatgtt agtactatcg attcacacaa aaaaccaaca cacagatgta
     7081 atgaaaataa agatatttta ttgatctagt aacatagatg acaccgcgcg cgataattta
     7141 tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact
     7201 ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
     7261 ttaacgtaat tcaacagaaa ttatatgata atcgtcgcaa gaccggcaac aggattcaat
     7321 c
//
