LOCUS       sequence-193323-        5787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U626 Guide acceptor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 5787)
  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: pICSL1P6_AtU626
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    447..468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(483..488,489..506,507..513)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    521..537
                     /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             555..566
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             573..1250
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1295..1366
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1382..1409
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        1423..1498
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(1510..1534)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1585..1609
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1765..2913)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3249..3837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3329..3348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4008..4799,4800..4868))
                     /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     4631..4650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4869..4973)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5060..5691
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    join(5783..5787,1..20)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt
       61 ttttaatgta ctggggtgga tgcagtgggc cccactctgt gaagacaatg tggaattcct
      121 cgagggagca tcttcattct taagatatga agataatctt caaaaggccc ctgggaatct
      181 gaaagaagag aagcaggccc atttatatgg gaaagaacaa tagtatttct tatataggcc
      241 catttaagtt gaaaacaatc ttcaaaagtc ccacatcgct tagataagaa aacgaagctg
      301 agtttatata cagctagagt cgaagtagtg attgtgagac ggatatcaat acgcaaaccg
      361 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      421 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      481 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      541 cacacatact agagaaagag gagaaatact agatggcttc ctccgaggac gttatcaaag
      601 agttcatgcg tttcaaagtt cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg
      661 aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac cgctaaactg aaagttacca
      721 aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc gcagttccag tacggttcca
      781 aagcttacgt taaacacccg gctgacatcc cggactacct gaaactgtcc ttcccggaag
      841 gtttcaaatg ggaacgtgtt atgaacttcg aggacggtgg tgttgttacc gttacccagg
      901 actcctccct gcaagacggt gagttcatct acaaagttaa actgcgtggt accaacttcc
      961 cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta
     1021 tgtacccgga ggacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg
     1081 gtggtcacta cgacgctgaa gttaaaacca cctacatggc taaaaaaccg gttcagctgc
     1141 cgggtgctta caaaaccgac atcaaactgg acatcacctc ccacaacgag gactacacca
     1201 tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac cggtgcttaa taacgctgat
     1261 agtgctagtg tagatcgcta ctagagccag gcatcaaata aaacgaaagg ctcagtcgaa
     1321 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac
     1381 tggctcacct tcgggtgggc ctttctgcgt ttatacgtct cagttttaga gctagaaata
     1441 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     1501 tttttctaga cccagctttc ttgtacaaag ttggcattac gctgagcttg tcttctgcac
     1561 gaagtggttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     1621 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     1681 atttgtatgt gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc
     1741 agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca
     1801 ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg
     1861 ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag
     1921 cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag
     1981 cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat
     2041 ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa
     2101 gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc
     2161 cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga
     2221 ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt
     2281 gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc
     2341 gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag
     2401 cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt
     2461 tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc
     2521 cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt
     2581 tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat
     2641 ggtcatcgac ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc
     2701 ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc
     2761 cgtagcttgc tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac
     2821 ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg
     2881 cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac
     2941 tcacgccggg gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag
     3001 ataatccacc ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta
     3061 cttggtattc cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc
     3121 gtgggcctcg gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt
     3181 gtcgccggca tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg
     3241 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3301 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3361 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3421 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3481 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3541 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3601 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3661 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3721 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3781 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3841 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3901 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3961 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4021 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4081 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4141 ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4201 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4261 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4321 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4381 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4441 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4501 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4561 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4621 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4681 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4741 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4801 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4861 atactcatac tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg
     4921 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat
     4981 tggccagcgc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg
     5041 gggatgggag gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc
     5101 ggcgtgcgcg gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt
     5161 ggtttaaaag caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg
     5221 caaatgctgg attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat
     5281 ctgtcagcac tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg
     5341 cccctcaagt gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg
     5401 aaactcgcgt aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc
     5461 ggccgaaatc gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa
     5521 gtgtcaacgt ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa
     5581 cgccggcggc cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg
     5641 ccatagacgg ccgccagccc agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag
     5701 atcctgatct gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
     5761 cggggagctg ttggctggct ggtggca
//
