LOCUS       sequence-193319-        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: pICSL1P4_AtU626
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      386..974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     875..894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1310..2458
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    2614..2638
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    2689..2713
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(2725..2800)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(2814..2841)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2857..2928)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2973..3650)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             3657..3668
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3686..3702
                     /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        complement(join(3710..3716,3717..3734,3735..3740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3755..3776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    4203..4227
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      4319..4950
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5037..5141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5142..5210,5211..5787,1..215)
                     /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(5360..5379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
       61 gtgagcgtgg ttctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      121 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
      181 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
      241 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
      301 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
      361 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      421 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      481 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      541 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      601 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      661 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      721 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      781 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      841 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      901 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      961 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggca gatcctagat
     1021 gtggcgcaac gatgccggcg acaagcagga gcgcaccgac ttcttccgca tcaagtgttt
     1081 tggctctcag gccgaggccc acggcaagta tttgggcaag gggtcgctgg tattcgtgca
     1141 gggcaagatt cggaatacca agtacgagaa ggacggccag acggtctacg ggaccgactt
     1201 cattgccgat aaggtggatt atctggacac caaggcacca ggcgggtcaa atcaggaata
     1261 agggcacatt gccccggcgt gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac
     1321 gtttgaccgg aaggcataca ggcaagaact gatcgacgcg gggttttccg ccgaggatgc
     1381 cgaaaccatc gcaagccgca ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg
     1441 ctcgatggtc cagcaagcta cggccaagat cgagcgcgac agcgtgcaac tggctccccc
     1501 tgccctgccc gcgccatcgg ccgccgtgga gcgttcgcgt cgtcttgaac aggaggcggc
     1561 aggtttggcg aagtcgatga ccatcgacac gcgaggaact atgacgacca agaagcgaaa
     1621 aaccgccggc gaggacctgg caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa
     1681 acacacgaag cagcagatca aggaaatgca gctttccttg ttcgatattg cgccgtggcc
     1741 ggacacgatg cgagcgatgc caaacgacac ggcccgctct gccctgttca ccacgcgcaa
     1801 caagaaaatc ccgcgcgagg cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt
     1861 gaagatcacc tacaccggcg tcgagctgcg ggccgacgat gacgaactgg tgtggcagca
     1921 ggtgttggag tacgcgaagc gcacccctat cggcgagccg atcaccttca cgttctacga
     1981 gctttgccag gacctgggct ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg
     2041 cctgtcgcgc ctacaggcga cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga
     2101 atcggtgtcg ctgctgcacc gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg
     2161 ccaggtcctg atcgacgagg aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt
     2221 catatgggag aagtaccgca agctgtcgcc gacggcccga cggatgttcg actatttcag
     2281 ctcgcaccgg gagccgtacc cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga
     2341 ttccacccgc gtgaagaagt ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg
     2401 cagcggcctg gtggaacacg cctgggtcaa tgatgacctg gtgcattgca aacgctaggg
     2461 ccttgtgggg tcagttccgg ctgggggttc agcagccagc gcctgatctg gggaaccctg
     2521 tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat
     2581 ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata tcctgtcaaa
     2641 cactgatagt ttaaaccact tcgtgcagaa gacaatctga gcgtaatgcc aactttgtac
     2701 aagaaagctg ggtctagaaa aaaagcaccg actcggtgcc actttttcaa gttgataacg
     2761 gactagcctt attttaactt gctatttcta gctctaaaac tgagacgtat aaacgcagaa
     2821 aggcccaccc gaaggtgagc cagtgtgact ctagtagaga gcgttcaccg acaaacaaca
     2881 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggctct
     2941 agtagcgatc tacactagca ctatcagcgt tattaagcac cggtggagtg acgaccttca
     3001 gcacgttcgt actgttcaac gatggtgtag tcctcgttgt gggaggtgat gtccagtttg
     3061 atgtcggttt tgtaagcacc cggcagctga accggttttt tagccatgta ggtggtttta
     3121 acttcagcgt cgtagtgacc accgtctttc agtttcagac gcattttgat ttcacctttc
     3181 agagcaccgt cctccgggta catacgttcg gtggaagctt cccaacccat ggtttttttc
     3241 tgcataaccg gaccgtcgga cgggaagttg gtaccacgca gtttaacttt gtagatgaac
     3301 tcaccgtctt gcagggagga gtcctgggta acggtaacaa caccaccgtc ctcgaagttc
     3361 ataacacgtt cccatttgaa accttccggg aaggacagtt tcaggtagtc cgggatgtca
     3421 gccgggtgtt taacgtaagc tttggaaccg tactggaact gcggggacag gatgtcccaa
     3481 gcgaacggca gcggaccacc tttggtaact ttcagtttag cggtctgggt accttcgtac
     3541 ggacgacctt caccttcacc ttcgatttcg aactcgtgac cgttaacgga accttccata
     3601 cgaactttga aacgcatgaa ctctttgata acgtcctcgg aggaagccat ctagtatttc
     3661 tcctctttct ctagtatgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3841 tcggccaacg cgcggggaga ggcggtttgc gtattgatat ccgtctcaca atcactactt
     3901 cgactctagc tgtatataaa ctcagcttcg ttttcttatc taagcgatgt gggacttttg
     3961 aagattgttt tcaacttaaa tgggcctata taagaaatac tattgttctt tcccatataa
     4021 atgggcctgc ttctcttctt tcagattccc aggggccttt tgaagattat cttcatatct
     4081 taagaatgaa gatgctcccc atgggaattc gtaattgtct tcacagagtg gggcccactg
     4141 catccacccc agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat
     4201 ttgtttacac cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc
     4261 ggcacaaaat caccactcga tacaggcagc ccatcagtca gatcaggatc tcctttgcga
     4321 cgctcaccgg gctggttgcc ctcgccgctg ggctggcggc cgtctatggc cctgcaaacg
     4381 cgccagaaac gccgtcgaag ccgtgtgcga gacaccgcgg ccgccggcgt tgtggatacc
     4441 tcgcggaaaa cttggccctc actgacagat gaggggcgga cgttgacact tgaggggccg
     4501 actcacccgg cgcggcgttg acagatgagg ggcaggctcg atttcggccg gcgacgtgga
     4561 gctggccagc ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc ccacagatga
     4621 tgtggacaag cctggggata agtgccctgc ggtattgaca cttgaggggc gcgactactg
     4681 acagatgagg ggcgcgatcc ttgacacttg aggggcagag tgctgacaga tgaggggcgc
     4741 acctattgac atttgagggg ctgtccacag gcagaaaatc cagcatttgc aagggtttcc
     4801 gcccgttttt cggccaccgc taacctgtct tttaacctgc ttttaaacca atatttataa
     4861 accttgtttt taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg
     4921 cccccccttc tcgaaccctc ccggcccgct aacgcgggcc tcccatcccc ccaggggctg
     4981 cgcccctcgg ccgcgaacgg cctcacccca aaaatggcag cgctggccaa ttcgtgcgcg
     5041 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5101 aaccttgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5161 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5221 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5281 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5341 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5401 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5461 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5521 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5581 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5641 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5701 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5761 actggatgga ggcggataaa gttgcag
//
