LOCUS       sequence-193317-        6532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GG compatible single transcript acceptor BASTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6532)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 6532)
  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: pICSL86955OD
FEATURES             Location/Qualifiers
     source          1..6532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(597..1226)
                     /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    1535..1559
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1719..2042)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1724..1743
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1937..1959
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1951..1968
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1952..1968
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1969..2025
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2038..2054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2038..2054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2051..2073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2062..2078
                     /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(2086..2092,2093..2110,2111..2116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2131..2152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      complement(2258..2510)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3110..3292)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(3146..3324)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    3487..3511
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3647..4235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3727..3746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4406..5200)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(5201..5305)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5367..5561
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(5627..6532,1..168))
                     /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 ggattctttg cggattcgat cagcggccgc ttgccacgat tcaccggggc gtgcttctgc
       61 ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac ttctccacca ggtcatcacc
      121 cagcgccgcg ccgatttgta ccgggccgga tggtttgcga ccgctcacgc cgattcctcg
      181 ggcttggggg ttccagtgcc attgcagggc cggcagacaa cccagccgct tacgcctggc
      241 caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg ttgttcttga
      301 ttttccatgc cgcctccttt agccgctaaa attcatctac tcatttattc atttgctcat
      361 ttactctggt agctgcgcga tgtattcaga tagcagctcg gtaatggtct tgccttggcg
      421 taccgcgtac atcttcagct tggtgtgatc ctccgccggc aactgaaagt tgacccgctt
      481 catggctggc gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc gtgcgctcgg
      541 acggccggca cttagcgtgt ttgtgctttt gctcattttc tctttacctc attaactcaa
      601 atgagttttg atttaatttc agcggccagc gcctggacct cgcgggcagc gtcgccctcg
      661 ggttctgatt caagaacggt tgtgccggcg gcggcagtgc ctgggtagct cacgcgctgc
      721 gtgatacggg actcaagaat gggcagctcg tacccggcca gcgcctcggc aacctcaccg
      781 ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag ccttccatcc
      841 gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg cggtggccca tatgtcgtaa
      901 gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga cacagccaag
      961 tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat ggccttcacg
     1021 tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc ttcccgcacg
     1081 gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac atcggccccg
     1141 gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga cccgcctttc
     1201 tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta tttactcatc
     1261 gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca catcaccttt
     1321 ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc gccagcttgg
     1381 catcagacaa accggccagg atttcatgca gccgcacggc tgactcccga ggggaaccct
     1441 gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc cttttaaata
     1501 tccgattatt ctaataaacg ctcttttctc ttaggtttac ccgccaatat atcctgtcaa
     1561 acactgatag tttaaacgga tcacttcgtg tccctcggtc acatgtgcat cctctagtag
     1621 cgtgagaccg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1681 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     1741 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1801 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     1861 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     1921 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
     1981 ggtacccggg gatcctctag agtcgacctg caggcatgca agcttggcgt aatcatggtc
     2041 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2101 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2161 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcggt ctcactccaa
     2221 gcttgaattc ttgcagcgct ccaattcagc aagcgtcgat ctagtaacat agatgacacc
     2281 gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg
     2341 tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg
     2401 ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat cgcaagaccg
     2461 gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttgactc
     2521 tagatccaga gtcccgctca aatctcagtc acaggaagaa caggtctagg aggaacagga
     2581 agagagaaat caagctgcca gaatcccaca tcatgccaat ttccatgttt gaaaccagca
     2641 gctctaagca ttcccctagg agcataacca agagcctcat gcatcctcac agaaggatca
     2701 ttaggaagtc caatcacagc aaccacagac ttgaaaccct gagcctccaa agacttcaaa
     2761 agatgagtgt aaagagtaga tccaagacca gttctttgat gcctaggaga cacgtaaaca
     2821 gtagactcag cagtccaatc gtaagcattc ctagccttcc aaggaccagc ataagcaata
     2881 ccagccactt caccatccac ttcagcaaca agccaaggat acctctctct aagcctcaca
     2941 agatcatcag tccactcttg tggttcttga ggctcagtcc tgaaattcac agtagaagtc
     3001 tcaatgtaat gattcacaat agtgcacaca gcaggcatat cagcctcagt agcccttcta
     3061 atatcagcag gccttctttc aggagacatg cgaaacgatc cagatccggt gcagattatt
     3121 tggattgaga gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag
     3181 tggagcattt tgacaagaaa tatttgctag ctgatagtga ccttaggcga cttttgaacg
     3241 cgcaataatg gtttctgacg tatgtgctta gctcattaaa ctccagaaac ccgcggctga
     3301 gtggctcctt caacgttgcg gttctgtcag ttccaaacgt aaaacggctt gtcccgcgtc
     3361 atcggcgggg gtcataacgt gactccctta attctccgct ccggatccga attcggcaca
     3421 gagtgttcaa ccccagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt
     3481 caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca
     3541 gctcggcaca aaatcaccac tcgatacagg cagcccatca accatgacta ctgtccggtc
     3601 caacagatcc gagcaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3661 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3721 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3781 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3841 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3901 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3961 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4021 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4081 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4141 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4261 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4381 tatatgagta aacttggtct gacagctaaa acaattcatc cagtaaaata taatatttta
     4441 ttttctccca atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt
     4501 ccccgatatc ctccctgagc gaccggacgc agaaggcaat gtcataccac ttgtccgccc
     4561 tgccgcttct cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc
     4621 tgtctcccag gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa
     4681 aatcatacag ctcgcgcgga tctttaaatg gagtgtcctc ttcccagttt tcgcaatcca
     4741 catcggccag atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat
     4801 tcgtataggg acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca
     4861 gctcgataat cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat
     4921 cggcctcact catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg
     4981 gaacaggcag ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg
     5041 tggtcccttt ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca
     5101 gcttatatac cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga
     5161 tcagtttttt caattccggt gatattctca ttttagccat actcttcctt tttcaatatt
     5221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5281 aaaataaaca aataggggtt ccgcgcactc ggagccttga tgtgggcgct ttttgcagct
     5341 gttcggctgt gcgctggcca gacagttatg cacaggccag gcgggtttta agagttttaa
     5401 taagttttaa agagttttag gcggaaaaat cgcctttttt ctcttttata tcagtcactt
     5461 acatgtgtga ccggttccca atgtacggct ttgggttccc aatgtacggg ttccggttcc
     5521 caatgtacgg ctttgggttc ccaatgtacg tgctatccac aggaaagaag ctttttcgac
     5581 ctttttcccc tgctagggca atttgcccta gcatctgctc cgtacattag gaaccggcgg
     5641 atgcttcgcc ctcgatcagg ttgcggtagc gcatgactag gatcgggcca gcctgccccg
     5701 cctcctcctt caaatcgtac tccggcaggt catttgaccc gatcagcttg cgcacggtga
     5761 aacagaactt cttgaactct ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc
     5821 atctggcttc tgccttgcct gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc
     5881 cgggatcgat caaaaagtaa tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg
     5941 tgatctcgcg gtacatccaa tcagctagct cgatctcgat gtactccggc cgcccggttt
     6001 cgctctttac gatcttgtag cggctaatca aggcttcacc ctcggatacc gtcaccaggc
     6061 ggccgttctt ggccttcttc gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc
     6121 aggtttctac caggtcgtct ttctgctttc cgccatcggc tcgccggcag aacttgagta
     6181 cgtccgcaac gtgtggacgg aacacgcggc cgggcttgtc tcccttccct tcccggtatc
     6241 ggttcatgga ttcggttaga tgggaaaccg ccatcagtac caggtcgtaa tcccacacac
     6301 tggccatgcc ggccggccct gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga
     6361 tcacctcgcc agctcgtcgg tcacgcttcg acagacggaa aacggccacg tccatgatgc
     6421 tgcgactatc gcgggtgccc acgtcataga gcatcggaac gaaaaaatct ggttgctcgt
     6481 cgcccttggg cggcttccta atcgacgacg caccggctgc cggcggttgc cg
//
