LOCUS       sequence-193593-        6671 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 6671)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 6671)
  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: pICSLUS0002OD
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    121..145
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        308..487
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    518..572
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             574..1368
                     /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(628..647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1238..1257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1395..2102
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    2212..2236
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2545..3174
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3603..4676
                     /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      4742..4936
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        4998..5102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5103..5897
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      6068..6656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6557..6576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctgct cggatctgtt ggaccggaca gtagtcatgg ttgatgggct
       61 gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg
      121 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
      181 gacgttttta atgtactggg gttgaacact ctgtgccgaa ttcggatccg gagcggagaa
      241 ttaagggagt cacgttatga cccccgccga tgacgcggga caagccgttt tacgtttgga
      301 actgacagaa ccgcaacgtt gaaggagcca ctgagccgcg ggtttctgga gtttaatgag
      361 ctaagcacat acgtcagaaa ccattattgc gcgttcaaaa gtcgcctaag gtcactatca
      421 gctagcaaat atttcttgtc aaaaatgctc cactgacgtt ccataaattc ccctcggtat
      481 ccaattagag tctcatattc actctcctat ttttacaaca attaccaaca acaacaaaca
      541 acaaacaaca ttacaattac atttacaatt accatggttg aacaagatgg attgcacgca
      601 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      661 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      721 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
      781 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      841 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      901 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      961 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     1021 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1081 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gactcatggc
     1141 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1201 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1261 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1321 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     1381 ggttcgctag agtcctgctt taatgagata tgcgagacgc ctatgatcgc atgatatttg
     1441 ctttcaattc tgttgtgcac gttgtaaaaa acctgagcat gtgtagctca gatccttacc
     1501 gccggtttcg gttcattcta atgaatatat cacccgttac tatcgtattt ttatgaataa
     1561 tattctccgt tcaatttact gattgtaccc tactacttat atgtacaata ttaaaatgaa
     1621 aacaatatat tgtgctgaat aggtttatag cgacatctat gatagagcgc cacaataaca
     1681 aacaattgcg ttttattatt acaaatccaa ttttaaaaaa agcggcagaa ccggtcaaac
     1741 ctaaaagact gattacataa atcttattca aatttcaaaa gtgccccagg ggctagtatc
     1801 tacgacacac cgagcggcga actaataacg ctcactgaag ggaactccgg ttccccgccg
     1861 gcgcgcatgg gtgagattcc ttgaagttga gtattggccg tccgctctac cgaaagttac
     1921 gggcaccatt caacccggtc cagcacggcg gccgggtaac cgacttgctg ccccgagaat
     1981 tatgcagcat ttttttggtg tatgtgggcc ccaaatgaag tgcaggtcaa accttgacag
     2041 tgacgacaaa tcgttgggcg ggtccagggc gaattttgcg acaacatgtc gaggctcagc
     2101 cgctgcaaga attcaagctt ggaggctgag gcttaattaa acctcagccg ctactagagg
     2161 atgcacatgt gaccgaggga cacgaagtga tccgtttaaa ctatcagtgt ttgacaggat
     2221 atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag
     2281 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc
     2341 tcgggagtca gccgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc
     2401 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg
     2461 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa
     2521 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg
     2581 tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat
     2641 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa
     2701 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc
     2761 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg
     2821 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata
     2881 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
     2941 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
     3001 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
     3061 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
     3121 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
     3181 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
     3241 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
     3301 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
     3361 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
     3421 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
     3481 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
     3541 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
     3601 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
     3661 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
     3721 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
     3781 cggcagccgg tgcgtcgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
     3841 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
     3901 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
     3961 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
     4021 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
     4081 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
     4141 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
     4201 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
     4261 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
     4321 acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg
     4381 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
     4441 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
     4501 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
     4561 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
     4621 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
     4681 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa agcttctttc
     4741 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
     4801 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
     4861 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
     4921 gcctggcctg tgcataactg tctggccagc gcacagccga acagctgcaa aaagcgccca
     4981 catcaaggct ccgagtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5041 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5101 gtatggctaa aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg
     5161 taaaagatac ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa
     5221 acctatattt aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg
     5281 aaaaggacat gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg
     5341 aacggcatga tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg
     5401 aagagtatga agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca
     5461 ggctctttca ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct
     5521 tagccgaatt ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg
     5581 aagaggacac tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc
     5641 ccgaagagga acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag
     5701 atggcaaagt aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg
     5761 acattgcctt ctgcgtccgg tcgctcaggg aggatatcgg ggaagaacag tatgtcgagc
     5821 tattttttga cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac
     5881 tggatgaatt gttttagctg tcagaccaag tttactcata tatactttag attgatttaa
     5941 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6001 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6061 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6121 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6181 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6241 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6301 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6361 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6421 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6481 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6541 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6601 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6661 ccagcaacgc g
//
