LOCUS       sequence-193314-        8678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GG compatible CDs acceptor BASTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8678)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 8678)
  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: pICSL86977OD
FEATURES             Location/Qualifiers
     source          1..8678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(689..1318)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    1627..1651
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1718..2425)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(2529..2852)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2534..2553
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2747..2769
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2761..2778
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2762..2778
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2779..2835
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2848..2864)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2848..2864)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2861..2883)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2872..2888
                     /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(2896..2902,2903..2920,2921..2926))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2941..2962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(3118..3463)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3155..3175)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4496..4748)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5348..5530)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5384..5562)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5725..5749
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5885..6473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5965..5984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6644..7438)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7439..7543)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7605..7799
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(7865..8678,1..260))
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 acgtcataga gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta
       61 atcgacgacg caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc
      121 gcttgccacg attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc
      181 gcggccttca acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg
      241 gatggtttgc gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg
      301 gccggcagac aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc
      361 attccacggc gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta
      421 aaattcatct actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca
      481 gatagcagct cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga
      541 tcctccgccg gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc
      601 aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt
      661 ttgctcattt tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca
      721 gcgcctggac ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg
      781 cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct
      841 cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg
      901 acacgacaaa ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct
      961 ccaccaggtc ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga atcagcacga
     1021 agtcggctgc cttgatcgcg gacacagcca agtccgccgc ctggggcgct ccgtcgatca
     1081 ctacgaagtc gcgccggccg atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc
     1141 cgacaacggt tagcggttga tcttcccgca cggccgccca atcgcgggca ctgccctggg
     1201 gatcggaatc gactaacaga acatcggccc cggcgagttg cagggcgcgg gctagatggg
     1261 ttgcgatggt cgtcttgcct gacccgcctt tctggttaag tacagcgata accttcatgc
     1321 gttccccttg cgtatttgtt tatttactca tcgcatcata tacgcagcga ccgcatgacg
     1381 caagctgttt tactcaaata cacatcacct ttttagacgg cggcgctcgg tttcttcagc
     1441 ggccaagctg gccggccagg ccgccagctt ggcatcagac aaaccggcca ggatttcatg
     1501 cagccgcacg gctgactccc gaggggaacc ctgtggttgg catgcacata caaatggacg
     1561 aacggataaa ccttttcacg cccttttaaa tatccgatta ttctaataaa cgctcttttc
     1621 tcttaggttt acccgccaat atatcctgtc aaacactgat agtttaaacg gatcacttcg
     1681 tgtccctcgg tcacatgtgc atcctctagt agcggtcctg ctgagcctcg acatgttgtc
     1741 gcaaaattcg ccctggaccc gcccaacgat ttgtcgtcac tgtcaaggtt tgacctgcac
     1801 ttcatttggg gcccacatac accaaaaaaa tgctgcataa ttctcggggc agcaagtcgg
     1861 ttacccggcc gccgtgctgg accgggttga atggtgcccg taactttcgg tagagcggac
     1921 ggccaatact caacttcaag gaatctcacc catgcgcgcc ggcggggaac cggagttccc
     1981 ttcagtgagc gttattagtt cgccgctcgg tgtgtcgtag atactagccc ctggggcact
     2041 tttgaaattt gaataagatt tatgtaatca gtcttttagg tttgaccggt tctgccgctt
     2101 tttttaaaat tggatttgta ataataaaac gcaattgttt gttattgtgg cgctctatca
     2161 tagatgtcgc tataaaccta ttcagcacaa tatattgttt tcattttaat attgtacata
     2221 taagtagtag ggtacaatca gtaaattgaa cggagaatat tattcataaa aatacgatag
     2281 taacgggtga tatattcatt agaatgaacc gaaaccggcg gtaaggatct gagctacaca
     2341 tgctcaggtt ttttacaacg tgcacaacag aattgaaagc aaatatcatg cgatcatagg
     2401 cttctcgcat atctcattaa agcaggacaa gctgagaccg tcacagcttg tctgtaagcg
     2461 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2521 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     2581 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     2641 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     2701 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     2761 tgtaaaacga cggccagtga attcgagctc ggtacccggg gatcctctag agtcgacctg
     2821 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2881 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2941 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3001 cctgtcgtgc cagctgcggt ctcccattgt atcgataatt gtaaatgtaa ttgtaatgtt
     3061 gtttgttgtt tgttgttgtt ggtaattgtt gtaaaaatga gctcttatac tcgagcgtgt
     3121 cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga
     3181 ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct ttgtagacgt
     3241 ggttggaacc tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc
     3301 actgtcggca gagagatctt gaatgatagc ctttccttta tcgcaatgat ggcatttgta
     3361 ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg ggcaatggaa
     3421 tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc tttgatcttc
     3481 tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca ccatgttgac
     3541 gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt cgccagtctt
     3601 gacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg gccttagatt
     3661 cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt ttatgaagca
     3721 agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg tctttctctg
     3781 tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc ttgggaaggt
     3841 atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagactt gctgcgtagg
     3901 cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta accttctcat
     3961 tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct agatcgtaaa
     4021 gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt ggggctggat
     4081 cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt ggtgggcttg
     4141 ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg atgaagttca
     4201 acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc tctgtaatag
     4261 tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt gtacaacccc
     4321 agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg atatgagggt
     4381 gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca gatttttgtg
     4441 ggattggaat tcctccaagc ttgaattctt gcagcgctcc aattcagcaa gcgtcgatct
     4501 agtaacatag atgacaccgc gcgcgataat ttatcctagt ttgcgcgcta tattttgttt
     4561 tctatcgcgt attaaatgta taattgcggg actctaatca taaaaaccca tctcataaat
     4621 aacgtcatgc attacatgtt aattattaca tgcttaacgt aattcaacag aaattatatg
     4681 ataatcatcg caagaccggc aacaggattc aatcttaaga aactttattg ccaaatgttt
     4741 gaacgatctg cttgactcta gatccagagt cccgctcaaa tctcagtcac aggaagaaca
     4801 ggtctaggag gaacaggaag agagaaatca agctgccaga atcccacatc atgccaattt
     4861 ccatgtttga aaccagcagc tctaagcatt cccctaggag cataaccaag agcctcatgc
     4921 atcctcacag aaggatcatt aggaagtcca atcacagcaa ccacagactt gaaaccctga
     4981 gcctccaaag acttcaaaag atgagtgtaa agagtagatc caagaccagt tctttgatgc
     5041 ctaggagaca cgtaaacagt agactcagca gtccaatcgt aagcattcct agccttccaa
     5101 ggaccagcat aagcaatacc agccacttca ccatccactt cagcaacaag ccaaggatac
     5161 ctctctctaa gcctcacaag atcatcagtc cactcttgtg gttcttgagg ctcagtcctg
     5221 aaattcacag tagaagtctc aatgtaatga ttcacaatag tgcacacagc aggcatatca
     5281 gcctcagtag cccttctaat atcagcaggc cttctttcag gagacatgcg aaacgatcca
     5341 gatccggtgc agattatttg gattgagagt gaatatgaga ctctaattgg ataccgaggg
     5401 gaatttatgg aacgtcagtg gagcattttg acaagaaata tttgctagct gatagtgacc
     5461 ttaggcgact tttgaacgcg caataatggt ttctgacgta tgtgcttagc tcattaaact
     5521 ccagaaaccc gcggctgagt ggctccttca acgttgcggt tctgtcagtt ccaaacgtaa
     5581 aacggcttgt cccgcgtcat cggcgggggt cataacgtga ctcccttaat tctccgctcc
     5641 ggatccgaat tcggcacaga gtgttcaacc ccagtacatt aaaaacgtcc gcaatgtgtt
     5701 attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca
     5761 acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcaac
     5821 catgactact gtccggtcca acagatccga gcaggaaccg taaaaaggcc gcgttgctgg
     5881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6481 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6541 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6601 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctaaaac aattcatcca
     6661 gtaaaatata atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata
     6721 gctcgacata ctgttcttcc ccgatatcct ccctgagcga ccggacgcag aaggcaatgt
     6781 cataccactt gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat
     6841 ctttcacaaa gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg
     6901 gcttttccgt ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcctctt
     6961 cccagttttc gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta
     7021 agcggctgtc taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc
     7081 tgatgcactc cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt
     7141 ccgagcaaag gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt
     7201 caaagtgcag gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt
     7261 cccgttccac atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt
     7321 tttcattttc tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta
     7381 cgcagcggta tttttcgatc agttttttca attccggtga tattctcatt ttagccatac
     7441 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7501 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcactcgg agccttgatg
     7561 tgggcgcttt ttgcagctgt tcggctgtgc gctggccaga cagttatgca caggccaggc
     7621 gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg ccttttttct
     7681 cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt gggttcccaa
     7741 tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg ctatccacag
     7801 gaaagaagct ttttcgacct ttttcccctg ctagggcaat ttgccctagc atctgctccg
     7861 tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc atgactagga
     7921 tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca tttgacccga
     7981 tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt
     8041 agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt
     8101 agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt
     8161 ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg atctcgatgt
     8221 actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag gcttcaccct
     8281 cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg
     8341 tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg ccatcggctc
     8401 gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc
     8461 ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc atcagtacca
     8521 ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct acgtgcccgt
     8581 ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac agacggaaaa
     8641 cggccacgtc catgatgctg cgactatcgc gggtgccc
//
