LOCUS       sequence-193595-        5964 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 5964)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 5964)
  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: pICSLUS0001OD
FEATURES             Location/Qualifiers
     source          1..5964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             126..1199
                     /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      1265..1459
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1521..1625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1626..2420
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2591..3179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3080..3099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3315..3339
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3502..3680
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        3534..3716
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      4316..4568
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4699..4723
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5032..5661
                     /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 caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc
       61 gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa
      121 tcggcgtgag cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg
      181 acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag
      241 cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac
      301 cgccggcagc cggtgcgtcg tcgattagga agccgcccaa gggcgacgag caaccagatt
      361 ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg
      421 ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc
      481 cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg
      541 acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga
      601 agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc
      661 ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca
      721 ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat
      781 ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg
      841 agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc
      901 cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc
      961 tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga
     1021 tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc
     1081 tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc
     1141 cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat
     1201 gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaagcttct
     1261 ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt
     1321 acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa
     1381 aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa
     1441 cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaacagctg caaaaagcgc
     1501 ccacatcaag gctccgagtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1561 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1621 agagtatggc taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct
     1681 gcgtaaaaga tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg
     1741 aaaacctata tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac
     1801 gggaaaagga catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact
     1861 ttgaacggca tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct
     1921 cggaagagta tgaagatgaa caaagccctg aaaagattat cgagctgtat gcggagtgca
     1981 tcaggctctt tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc
     2041 gcttagccga attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact
     2101 gggaagagga cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa
     2161 agcccgaaga ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga
     2221 aagatggcaa agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt
     2281 atgacattgc cttctgcgtc cggtcgctca gggaggatat cggggaagaa cagtatgtcg
     2341 agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt
     2401 tactggatga attgttttag ctgtcagacc aagtttactc atatatactt tagattgatt
     2461 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2521 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2581 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2641 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2701 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2761 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2821 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2881 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2941 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3001 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3061 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3121 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3181 acgccagcaa cgcggccttt ttacggttcc tgctcggatc tgttggaccg gacagtagtc
     3241 atggttgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
     3301 tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt
     3361 aataacacat tgcggacgtt tttaatgtac tggggttgaa cactctgtgc cgaattcgga
     3421 tccggagcgg agaattaagg gagtcacgtt atgacccccg ccgatgacgc gggacaagcc
     3481 gttttacgtt tggaactgac agaaccgcaa cgttgaagga gccactcagc cgcgggtttc
     3541 tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc
     3601 taaggtcact atcagctagc aaatatttct tgtcaaaatg ctccactgac gttccataaa
     3661 ttcccctcgg tatccaatta gagtctcata ttcactctca atccaaataa tctgcaccgg
     3721 atctggatcg tttcgcatgt ctcctgaaag aaggcctgct gatattagaa gggctactga
     3781 ggctgatatg cctgctgtgt gcactattgt gaatcattac attgagactt ctactgtgaa
     3841 tttcaggact gagcctcaag aaccacaaga gtggactgat gatcttgtga ggcttagaga
     3901 gaggtatcct tggcttgttg ctgaagtgga tggtgaagtg gctggtattg cttatgctgg
     3961 tccttggaag gctaggaatg cttacgattg gactgctgag tctactgttt acgtgtctcc
     4021 taggcatcaa agaactggtc ttggatctac tctttacact catcttttga agtctttgga
     4081 ggctcagggt ttcaagtctg tggttgctgt gattggactt cctaatgatc cttctgtgag
     4141 gatgcatgag gctcttggtt atgctcctag gggaatgctt agagctgctg gtttcaaaca
     4201 tggaaattgg catgatgtgg gattctggca gcttgatttc tctcttcctg ttcctcctag
     4261 acctgttctt cctgtgactg agatttgagc gggactctgg atctagagtc aagcagatcg
     4321 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     4381 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     4441 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     4501 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     4561 actagatcga cgcttgctga attggagcgc tgcaagaatt caagcttgga ggctgaggct
     4621 taattaaacc tcagccgcta ctagaggatg cacatgtgac cgagggacac gaagtgatcc
     4681 gtttaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag aaaagagcgt
     4741 ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt
     4801 atgtgcatgc caaccacagg gttcccctcg ggagtcagcc gtgcggctgc atgaaatcct
     4861 ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac
     4921 cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt
     4981 cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt
     5041 atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc
     5101 cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt
     5161 gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc
     5221 ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga
     5281 gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt
     5341 ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag
     5401 cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc
     5461 aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt
     5521 cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc
     5581 gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa
     5641 attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca
     5701 cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg
     5761 cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc
     5821 tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg
     5881 cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa
     5941 ggaggcggca tggaaaatca agaa
//
