LOCUS       sequence-193309-        7239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GG compatible single transcript acceptor NPTII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7239)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 7239)
  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: pICSL86900OD
FEATURES             Location/Qualifiers
     source          1..7239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(603..1232)
                     /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    1541..1565
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1725..2048)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1730..1749
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1943..1965
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1957..1974
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1958..1974
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1975..2031
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2044..2060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2044..2060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2057..2079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2068..2084
                     /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(2092..2098,2099..2116,2117..2122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2137..2158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      complement(2243..2950)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(2977..3771)
                     /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(3088..3107)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3698..3717
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_feature    3773..3827
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     promoter        complement(3858..4037)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4200..4224
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4360..4948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4440..4459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5119..5913)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(5914..6018)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6080..6274
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(6340..7239,1..174))
                     /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 ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc cacgattcac cggggcgtgc
       61 ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct ccaccaggtc
      121 atcacccagc gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc tcacgccgat
      181 tcctcgggct tgggggttcc agtgccattg cagggccggc agacaaccca gccgcttacg
      241 cctggccaac cgcccgttcc tccacacatg gggcattcca cggcgtcggt gcctggttgt
      301 tcttgatttt ccatgccgcc tcctttagcc gctaaaattc atctactcat ttattcattt
      361 gctcatttac tctggtagct gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc
      421 ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc gccggcaact gaaagttgac
      481 ccgcttcatg gctggcgtgt ctgccaggct ggccaatgtt gcagccttgc tgctgcgtgc
      541 gctcggacgg ccggcactta gcgtgtttgt gcttttgctc attttctctt tacctcatta
      601 actcaaatga gttttgattt aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg
      661 ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg
      721 cgctgcgtga tacgggactc aagaatgggc agctcgtacc cggccagcgc ctcggcaacc
      781 tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt
      841 ccatccgtga cctcaatgcg ctgcttaacc agctccacca ggtcggcggt ggcccatatg
      901 tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca
      961 gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc
     1021 ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc
     1081 cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg
     1141 gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg
     1201 cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta
     1261 ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc
     1321 acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca
     1381 gcttggcatc agacaaaccg gccaggattt catgcagccg cacggctgac tcccgagggg
     1441 aaccctgtgg ttggcatgca catacaaatg gacgaacgga taaacctttt cacgcccttt
     1501 taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc caatatatcc
     1561 tgtcaaacac tgatagttta aacggatcac ttcgtgtccc tcggtcacat gtgcatcctc
     1621 tagtagcgtg agaccgtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     1681 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     1741 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     1801 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1861 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     1921 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc
     1981 gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct tggcgtaatc
     2041 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2101 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2161 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcggtctca
     2221 ctccaagctt gaattcttgc agcggctgag cctcgacatg ttgtcgcaaa attcgccctg
     2281 gacccgccca acgatttgtc gtcactgtca aggtttgacc tgcacttcat ttggggccca
     2341 catacaccaa aaaaatgctg cataattctc ggggcagcaa gtcggttacc cggccgccgt
     2401 gctggaccgg gttgaatggt gcccgtaact ttcggtagag cggacggcca atactcaact
     2461 tcaaggaatc tcacccatgc gcgccggcgg ggaaccggag ttcccttcag tgagcgttat
     2521 tagttcgccg ctcggtgtgt cgtagatact agcccctggg gcacttttga aatttgaata
     2581 agatttatgt aatcagtctt ttaggtttga ccggttctgc cgcttttttt aaaattggat
     2641 ttgtaataat aaaacgcaat tgtttgttat tgtggcgctc tatcatagat gtcgctataa
     2701 acctattcag cacaatatat tgttttcatt ttaatattgt acatataagt agtagggtac
     2761 aatcagtaaa ttgaacggag aatattattc ataaaaatac gatagtaacg ggtgatatat
     2821 tcattagaat gaaccgaaac cggcggtaag gatctgagct acacatgctc aggtttttta
     2881 caacgtgcac aacagaattg aaagcaaata tcatgcgatc ataggcgtct cgcatatctc
     2941 attaaagcag gactctagcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     3001 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3061 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3121 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3181 catgatattc ggcaagcagg catcgccatg agtcacgacg agatcctcgc cgtcgggcat
     3241 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3301 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3361 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3421 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     3481 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     3541 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     3601 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     3661 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     3721 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaacca tggtaattgt
     3781 aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg taattgttgt aaaaatagga
     3841 gagtgaatat gagactctaa ttggataccg aggggaattt atggaacgtc agtggagcat
     3901 ttttgacaag aaatatttgc tagctgatag tgaccttagg cgacttttga acgcgcaata
     3961 atggtttctg acgtatgtgc ttagctcatt aaactccaga aacccgcggc tcagtggctc
     4021 cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg
     4081 ggggtcataa cgtgactccc ttaattctcc gctccggatc cgaattcggc acagagtgtt
     4141 caaccccagt acattaaaaa cgtccgcaat gtgttattaa gttgtctaag cgtcaatttg
     4201 tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg gcagctcggc
     4261 acaaaatcac cactcgatac aggcagccca tcaaccatga ctactgtccg gtccaacaga
     4321 tccgagcagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4801 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5101 gtaaacttgg tctgacagct aaaacaattc atccagtaaa atataatatt ttattttctc
     5161 ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat
     5221 atcctccctg agcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct
     5281 tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc
     5341 caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata
     5401 cagctcgcgc ggatctttaa atggagtgtc ctcttcccag ttttcgcaat ccacatcggc
     5461 cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata
     5521 gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat
     5581 aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc
     5641 actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg
     5701 cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc
     5761 tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata
     5821 taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt
     5881 tttcaattcc ggtgatattc tcattttagc catactcttc ctttttcaat attattgaag
     5941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6001 acaaataggg gttccgcgca ctcggagcct tgatgtgggc gctttttgca gctgttcggc
     6061 tgtgcgctgg ccagacagtt atgcacaggc caggcgggtt ttaagagttt taataagttt
     6121 taaagagttt taggcggaaa aatcgccttt tttctctttt atatcagtca cttacatgtg
     6181 tgaccggttc ccaatgtacg gctttgggtt cccaatgtac gggttccggt tcccaatgta
     6241 cggctttggg ttcccaatgt acgtgctatc cacaggaaag aagctttttc gacctttttc
     6301 ccctgctagg gcaatttgcc ctagcatctg ctccgtacat taggaaccgg cggatgcttc
     6361 gccctcgatc aggttgcggt agcgcatgac taggatcggg ccagcctgcc ccgcctcctc
     6421 cttcaaatcg tactccggca ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa
     6481 cttcttgaac tctccggcgc tgccactgcg ttcgtagatc gtcttgaaca accatctggc
     6541 ttctgccttg cctgcggcgc ggcgtgccag gcggtagaga aaacggccga tgccgggatc
     6601 gatcaaaaag taatcggggt gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc
     6661 gcggtacatc caatcagcta gctcgatctc gatgtactcc ggccgcccgg tttcgctctt
     6721 tacgatcttg tagcggctaa tcaaggcttc accctcggat accgtcacca ggcggccgtt
     6781 cttggccttc ttcgtacgct gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc
     6841 taccaggtcg tctttctgct ttccgccatc ggctcgccgg cagaacttga gtacgtccgc
     6901 aacgtgtgga cggaacacgc ggccgggctt gtctcccttc ccttcccggt atcggttcat
     6961 ggattcggtt agatgggaaa ccgccatcag taccaggtcg taatcccaca cactggccat
     7021 gccggccggc cctgcggaaa cctctacgtg cccgtctgga agctcgtagc ggatcacctc
     7081 gccagctcgt cggtcacgct tcgacagacg gaaaacggcc acgtccatga tgctgcgact
     7141 atcgcgggtg cccacgtcat agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt
     7201 gggcggcttc ctaatcgacg acgcaccggc tgccggcgg
//
