LOCUS       sequence-193589-        7196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GG compatible USER cloning .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 7196)
  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: pICSLUS0003OD
FEATURES             Location/Qualifiers
     source          1..7196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             69..1142
                     /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"
     rep_origin      1208..1402
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1464..1568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1569..2363
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2534..3122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3023..3042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3258..3282
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3445..3623
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        3477..3659
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4259..4511
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        4632..4977
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4920..4940
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      5100..5807
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    5874..5898
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6207..6836
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 ggcgtaagcg gctgggttgt ctgccggccc tgcaatggca ctggaacccc caagcccgag
       61 gaatcggcgt gagcggtcgc aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg
      121 atgacctggt ggagaagttg aaggccgcgc aggccgccca gcggcaacgc atcgaggcag
      181 aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc
      241 aaccgccggc agccggtgcg tcgtcgatta ggaagccgcc caagggcgac gagcaaccag
      301 attttttcgt tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc atcatggacg
      361 tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc
      421 ttccagacgg gcacgtagag gtttccgcag ggccggccgg catggccagt gtgtgggatt
      481 acgacctggt actgatggcg gtttcccatc taaccgaatc catgaaccga taccgggaag
      541 ggaagggaga caagcccggc cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct
      601 gccggcgagc cgatggcgga aagcagaaag acgacctggt agaaacctgc attcggttaa
      661 acaccacgca cgttgccatg cagcgtacga agaaggccaa gaacggccgc ctggtgacgg
      721 tatccgaggg tgaagccttg attagccgct acaagatcgt aaagagcgaa accgggcggc
      781 cggagtacat cgagatcgag ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga
      841 acccggacgt gctgacggtt caccccgatt actttttgat cgatcccggc atcggccgtt
      901 ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga
      961 cgatctacga acgcagtggc agcgccggag agttcaagaa gttctgtttc accgtgcgca
     1021 agctgatcgg gtcaaatgac ctgccggagt acgatttgaa ggaggaggcg gggcaggctg
     1081 gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg cgaagcatcc gccggttcct
     1141 aatgtacgga gcagatgcta gggcaaattg ccctagcagg ggaaaaaggt cgaaaaagct
     1201 tctttcctgt ggatagcacg tacattggga acccaaagcc gtacattggg aaccggaacc
     1261 cgtacattgg gaacccaaag ccgtacattg ggaaccggtc acacatgtaa gtgactgata
     1321 taaaagagaa aaaaggcgat ttttccgcct aaaactcttt aaaacttatt aaaactctta
     1381 aaacccgcct ggcctgtgca taactgtctg gccagcgcac agccgaacag ctgcaaaaag
     1441 cgcccacatc aaggctccga gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1501 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1561 ggaagagtat ggctaaaatg agaatatcac cggaattgaa aaaactgatc gaaaaatacc
     1621 gctgcgtaaa agatacggaa ggaatgtctc ctgctaaggt atataagctg gtgggagaaa
     1681 atgaaaacct atatttaaaa atgacggaca gccggtataa agggaccacc tatgatgtgg
     1741 aacgggaaaa ggacatgatg ctatggctgg aaggaaagct gcctgttcca aaggtcctgc
     1801 actttgaacg gcatgatggc tggagcaatc tgctcatgag tgaggccgat ggcgtccttt
     1861 gctcggaaga gtatgaagat gaacaaagcc ctgaaaagat tatcgagctg tatgcggagt
     1921 gcatcaggct ctttcactcc atcgacatat cggattgtcc ctatacgaat agcttagaca
     1981 gccgcttagc cgaattggat tacttactga ataacgatct ggccgatgtg gattgcgaaa
     2041 actgggaaga ggacactcca tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg
     2101 aaaagcccga agaggaactt gtcttttccc acggcgacct gggagacagc aacatctttg
     2161 tgaaagatgg caaagtaagt ggctttattg atcttgggag aagcggcagg gcggacaagt
     2221 ggtatgacat tgccttctgc gtccggtcgc tcagggagga tatcggggaa gaacagtatg
     2281 tcgagctatt ttttgactta ctggggatca agcctgattg ggagaaaata aaatattata
     2341 ttttactgga tgaattgttt tagctgtcag accaagttta ctcatatata ctttagattg
     2401 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2461 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2521 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2581 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3001 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3061 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3121 aaaacgccag caacgcggcc tttttacggt tcctgctcgg atctgttgga ccggacagta
     3181 gtcatggttg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg
     3241 agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa
     3301 cttaataaca cattgcggac gtttttaatg tactggggtt gaacactctg tgccgaattc
     3361 ggatccggag cggagaatta agggagtcac gttatgaccc ccgccgatga cgcgggacaa
     3421 gccgttttac gtttggaact gacagaaccg caacgttgaa ggagccactc agccgcgggt
     3481 ttctggagtt taatgagcta agcacatacg tcagaaacca ttattgcgcg ttcaaaagtc
     3541 gcctaaggtc actatcagct agcaaatatt tcttgtcaaa atgctccact gacgttccat
     3601 aaattcccct cggtatccaa ttagagtctc atattcactc tcaatccaaa taatctgcac
     3661 cggatctgga tcgtttcgca tgtctcctga aagaaggcct gctgatatta gaagggctac
     3721 tgaggctgat atgcctgctg tgtgcactat tgtgaatcat tacattgaga cttctactgt
     3781 gaatttcagg actgagcctc aagaaccaca agagtggact gatgatcttg tgaggcttag
     3841 agagaggtat ccttggcttg ttgctgaagt ggatggtgaa gtggctggta ttgcttatgc
     3901 tggtccttgg aaggctagga atgcttacga ttggactgct gagtctactg tttacgtgtc
     3961 tcctaggcat caaagaactg gtcttggatc tactctttac actcatcttt tgaagtcttt
     4021 ggaggctcag ggtttcaagt ctgtggttgc tgtgattgga cttcctaatg atccttctgt
     4081 gaggatgcat gaggctcttg gttatgctcc taggggaatg cttagagctg ctggtttcaa
     4141 acatggaaat tggcatgatg tgggattctg gcagcttgat ttctctcttc ctgttcctcc
     4201 tagacctgtt cttcctgtga ctgagatttg agcgggactc tggatctaga gtcaagcaga
     4261 tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat
     4321 gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat
     4381 gacgttattt atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc
     4441 gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat
     4501 gttactagat cgacgcttgc tgaattggag cgctgcaaga attcaagctt ggaggtcaac
     4561 atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt ctcagaagat
     4621 caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct cggattccat
     4681 tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa
     4741 tgccatcatt gcgataaagg aaaggctatc attcaagatc tctctgccga cagtggtccc
     4801 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagaggttcc aaccacgtct
     4861 acaaagcaag tggattgatg tgacatctcc actgacgtaa gggatgacgc acaatcccac
     4921 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacacgc
     4981 tcgagtataa gagctcattt ttacaacaat taccaacaac aacaaacaac aaacaacatt
     5041 acaattacat ttacaattat cgatacaagc tgaggcttaa ttaaacctca gcgcttgtcc
     5101 tgctttaatg agatatgcga gaagcctatg atcgcatgat atttgctttc aattctgttg
     5161 tgcacgttgt aaaaaacctg agcatgtgta gctcagatcc ttaccgccgg tttcggttca
     5221 ttctaatgaa tatatcaccc gttactatcg tatttttatg aataatattc tccgttcaat
     5281 ttactgattg taccctacta cttatatgta caatattaaa atgaaaacaa tatattgtgc
     5341 tgaataggtt tatagcgaca tctatgatag agcgccacaa taacaaacaa ttgcgtttta
     5401 ttattacaaa tccaatttta aaaaaagcgg cagaaccggt caaacctaaa agactgatta
     5461 cataaatctt attcaaattt caaaagtgcc ccaggggcta gtatctacga cacaccgagc
     5521 ggcgaactaa taacgctcac tgaagggaac tccggttccc cgccggcgcg catgggtgag
     5581 attccttgaa gttgagtatt ggccgtccgc tctaccgaaa gttacgggca ccattcaacc
     5641 cggtccagca cggcggccgg gtaaccgact tgctgccccg agaattatgc agcatttttt
     5701 tggtgtatgt gggccccaaa tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt
     5761 gggcgggtcc agggcgaatt ttgcgacaac atgtcgaggc tcagcaggac cgctactaga
     5821 ggatgcacat gtgaccgagg gacacgaagt gatccgttta aactatcagt gtttgacagg
     5881 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
     5941 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
     6001 cctcgggagt cagccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg
     6061 gcggcctggc cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga
     6121 tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta
     6181 aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg
     6241 ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg
     6301 atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg
     6361 aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg
     6421 ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg
     6481 tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca
     6541 tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa
     6601 ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg
     6661 ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg
     6721 tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg
     6781 acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta
     6841 atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc
     6901 acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt
     6961 caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
     7021 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
     7081 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
     7141 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggcca
//
