LOCUS       sequence-193591-        8052 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 8052)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 8052)
  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: pICSLUS0004OD
FEATURES             Location/Qualifiers
     source          1..8052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    36..60
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             369..998
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1427..2500
                     /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      2566..2760
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        2971..3075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3076..3870
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      4041..4629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4530..4549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4765..4789
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4952..5131
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5162..5216
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             5218..6012
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5272..5291)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5882..5901
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6039..6746
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        6846..7191
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     7134..7154
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      7314..8021
                     /label=OCS terminator
                     /note="octopine synthase terminator"
ORIGIN
        1 gggacacgaa gtgatccgtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
       61 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
      121 tccgttcgtc catttgtatg tgcatgccaa ccacagggtt cccctcggga gtcagccgtg
      181 cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc
      241 ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac
      301 agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag
      361 gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca
      421 tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt
      481 ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg
      541 ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact
      601 tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag
      661 ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc
      721 tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg
      781 tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg
      841 ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg
      901 ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc
      961 aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat
     1021 gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc
     1081 aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc
     1141 ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct
     1201 gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat
     1261 gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg
     1321 tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct
     1381 gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa
     1441 accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa
     1501 ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg
     1561 gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgtc
     1621 gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta
     1681 tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa
     1741 gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt
     1801 ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt
     1861 ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg
     1921 cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa
     1981 gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca
     2041 gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat
     2101 tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct
     2161 agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca
     2221 ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg
     2281 cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag
     2341 cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct
     2401 gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta
     2461 ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg
     2521 gcaaattgcc ctagcagggg aaaaaggtcg aaaaagcttc tttcctgtgg atagcacgta
     2581 cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc
     2641 gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt
     2701 ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata
     2761 actgtctggc cagcgcacag ccgaacagct gcaaaaagcg cccacatcaa ggctccgagt
     2821 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2881 acaatnanaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaacagctg
     2941 caaaaagcgc ccacatcaag gctccgagtg cgcggaaccc ctatttgttt atttttctaa
     3001 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3061 tgaaaaagga agagtatggc taaaatgaga atatcaccgg aattgaaaaa actgatcgaa
     3121 aaataccgct gcgtaaaaga tacggaagga atgtctcctg ctaaggtata taagctggtg
     3181 ggagaaaatg aaaacctata tttaaaaatg acggacagcc ggtataaagg gaccacctat
     3241 gatgtggaac gggaaaagga catgatgcta tggctggaag gaaagctgcc tgttccaaag
     3301 gtcctgcact ttgaacggca tgatggctgg agcaatctgc tcatgagtga ggccgatggc
     3361 gtcctttgct cggaagagta tgaagatgaa caaagccctg aaaagattat cgagctgtat
     3421 gcggagtgca tcaggctctt tcactccatc gacatatcgg attgtcccta tacgaatagc
     3481 ttagacagcc gcttagccga attggattac ttactgaata acgatctggc cgatgtggat
     3541 tgcgaaaact gggaagagga cactccattt aaagatccgc gcgagctgta tgatttttta
     3601 aagacggaaa agcccgaaga ggaacttgtc ttttcccacg gcgacctggg agacagcaac
     3661 atctttgtga aagatggcaa agtaagtggc tttattgatc ttgggagaag cggcagggcg
     3721 gacaagtggt atgacattgc cttctgcgtc cggtcgctca gggaggatat cggggaagaa
     3781 cagtatgtcg agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa
     3841 tattatattt tactggatga attgttttag ctgtcagacc aagtttactc atatatactt
     3901 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3961 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4021 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4081 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4141 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4201 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4261 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4321 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4381 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4441 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4501 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4561 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4621 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tgctcggatc tgttggaccg
     4681 gacagtagtc atggttgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     4741 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
     4801 tagacaactt aataacacat tgcggacgtt tttaatgtac tggggttgaa cactctgtgc
     4861 cgaattcgga tccggagcgg agaattaagg gagtcacgtt atgacccccg ccgatgacgc
     4921 gggacaagcc gttttacgtt tggaactgac agaaccgcaa cgttgaagga gccactgagc
     4981 cgcgggtttc tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc
     5041 aaaagtcgcc taaggtcact atcagctagc aaatatttct tgtcaaaaat gctccactga
     5101 cgttccataa attcccctcg gtatccaatt agagtctcat attcactctc ctatttttac
     5161 aacaattacc aacaacaaca aacaacaaac aacattacaa ttacatttac aattaccatg
     5221 gttgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5281 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5341 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5401 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5461 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5521 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5581 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5641 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5701 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5761 gaggatctcg tcgtgactca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5821 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5881 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5941 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6001 gagttcttct gagcgggact ctggggttcg ctagagtcct gctttaatga gatatgcgag
     6061 acgcctatga tcgcatgata tttgctttca attctgttgt gcacgttgta aaaaacctga
     6121 gcatgtgtag ctcagatcct taccgccggt ttcggttcat tctaatgaat atatcacccg
     6181 ttactatcgt atttttatga ataatattct ccgttcaatt tactgattgt accctactac
     6241 ttatatgtac aatattaaaa tgaaaacaat atattgtgct gaataggttt atagcgacat
     6301 ctatgataga gcgccacaat aacaaacaat tgcgttttat tattacaaat ccaattttaa
     6361 aaaaagcggc agaaccggtc aaacctaaaa gactgattac ataaatctta ttcaaatttc
     6421 aaaagtgccc caggggctag tatctacgac acaccgagcg gcgaactaat aacgctcact
     6481 gaagggaact ccggttcccc gccggcgcgc atgggtgaga ttccttgaag ttgagtattg
     6541 gccgtccgct ctaccgaaag ttacgggcac cattcaaccc ggtccagcac ggcggccggg
     6601 taaccgactt gctgccccga gaattatgca gcattttttt ggtgtatgtg ggccccaaat
     6661 gaagtgcagg tcaaaccttg acagtgacga caaatcgttg ggcgggtcca gggcgaattt
     6721 tgcgacaaca tgtcgaggct cagccgctgc aagaattcaa gcttggaggt caacatggtg
     6781 gagcacgaca ctctggtcta ctccaaaaat gtcaaagata cagtctcaga agatcaaagg
     6841 gctattgaga cttttcaaca aaggataatt tcgggaaacc tcctcggatt ccattgccca
     6901 gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta caaatgccat
     6961 cattgcgata aaggaaaggc tatcattcaa gatctctctg ccgacagtgg tcccaaagat
     7021 ggacccccac ccacgaggag catcgtggaa aaagaagagg ttccaaccac gtctacaaag
     7081 caagtggatt gatgtgacat ctccactgac gtaagggatg acgcacaatc ccactatcct
     7141 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac acgctcgagt
     7201 ataagagctc atttttacaa caattaccaa caacaacaaa caacaaacaa cattacaatt
     7261 acatttacaa ttatcgatac aagctgaggc ttaattaaac ctcagcgctt gtcctgcttt
     7321 aatgagatat gcgagaagcc tatgatcgca tgatatttgc tttcaattct gttgtgcacg
     7381 ttgtaaaaaa cctgagcatg tgtagctcag atccttaccg ccggtttcgg ttcattctaa
     7441 tgaatatatc acccgttact atcgtatttt tatgaataat attctccgtt caatttactg
     7501 attgtaccct actacttata tgtacaatat taaaatgaaa acaatatatt gtgctgaata
     7561 ggtttatagc gacatctatg atagagcgcc acaataacaa acaattgcgt tttattatta
     7621 caaatccaat tttaaaaaaa gcggcagaac cggtcaaacc taaaagactg attacataaa
     7681 tcttattcaa atttcaaaag tgccccaggg gctagtatct acgacacacc gagcggcgaa
     7741 ctaataacgc tcactgaagg gaactccggt tccccgccgg cgcgcatggg tgagattcct
     7801 tgaagttgag tattggccgt ccgctctacc gaaagttacg ggcaccattc aacccggtcc
     7861 agcacggcgg ccgggtaacc gacttgctgc cccgagaatt atgcagcatt tttttggtgt
     7921 atgtgggccc caaatgaagt gcaggtcaaa ccttgacagt gacgacaaat cgttgggcgg
     7981 gtccagggcg aattttgcga caacatgtcg aggctcagca ggaccgctac tagaggatgc
     8041 acatgtgacc ga
//
