LOCUS       sequence-192856-        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: pICSL1P1_AtU626
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..678
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      723..794
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      810..837
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        851..926
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(938..962)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1013..1037
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1193..2341)
                     /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(2677..3265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2757..2776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3436..4227,4228..4296))
                     /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     4059..4078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4297..4401)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4488..5119
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5211..5235
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    5662..5683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5698..5703,5704..5721,5722..5728)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5736..5752
                     /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             5770..5781
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atggcttcct ccgaggacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt
       61 tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt
      121 acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc
      181 ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg
      241 gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgag
      301 gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac
      361 aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc
      421 atgggttggg aagcttccac cgaacgtatg tacccggagg acggtgctct gaaaggtgaa
      481 atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc
      541 tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac
      601 atcacctccc acaacgagga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt
      661 cactccaccg gtgcttaata acgctgatag tgctagtgta gatcgctact agagccaggc
      721 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      781 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
      841 atacgtctca gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      901 ttgaaaaagt ggcaccgagt cggtgctttt tttctagacc cagctttctt gtacaaagtt
      961 ggcattacgc tgcaattgtc ttctgcacga agtggtttaa actatcagtg tttgacagga
     1021 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     1081 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     1141 cagatcaggc gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt
     1201 tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact
     1261 cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc
     1321 acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt
     1381 cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt
     1441 agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg
     1501 ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc
     1561 caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg
     1621 ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg
     1681 tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca
     1741 ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt
     1801 tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg
     1861 tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg
     1921 caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg
     1981 cggcctgctt ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct
     2041 tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct
     2101 gttcaagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca
     2161 gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact
     2221 ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg
     2281 cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca
     2341 ttcaccctcc ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat
     2401 ttgacccgcc tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc
     2461 cgtagaccgt ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata
     2521 ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga
     2581 tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc
     2641 tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3541 cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga
     3601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4321 aagcatttat caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4381 taaacaaata ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg
     4441 ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag
     4501 ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc
     4561 ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg
     4621 gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc
     4681 ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga
     4741 tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat
     4801 ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat
     4861 ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa
     4921 cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag
     4981 ggccaagttt tccgcgaggt atccacaacg ccggcggccg cggtgtctcg cacacggctt
     5041 cgacggcgtt tctggcgcgt ttgcagggcc atagacggcc gccagcccag cggcgagggc
     5101 aaccagcccg gtgagcgtcg caaaggagat cctgatctga ctgatgggct gcctgtatcg
     5161 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
     5221 tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta
     5281 atgtactggg gtggatgcag tgggccccac tctgtgaaga caatgccgaa ttcggatccg
     5341 gagcatcttc attcttaaga tatgaagata atcttcaaaa ggcccctggg aatctgaaag
     5401 aagagaagca ggcccattta tatgggaaag aacaatagta tttcttatat aggcccattt
     5461 aagttgaaaa caatcttcaa aagtcccaca tcgcttagat aagaaaacga agctgagttt
     5521 atatacagct agagtcgaag tagtgattgt gagacggata tcaatacgca aaccgcctct
     5581 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5641 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5701 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5761 atactagaga aagaggagaa atactag
//
