LOCUS       sequence-193313-        5313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LevelP cloning Vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  TITLE     Youles SynBio plasmids
REFERENCE   2  (bases 1 to 5313)
  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: pICSL4723-P1
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             157..1230
                     /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      1296..1490
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        1552..1656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1657..2451
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2622..3210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3111..3130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3346..3370
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    3515..3536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3551..3556,3557..3574,3575..3581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3589..3605
                     /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)."
     primer_bind     3594..3616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3613..3629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3613..3629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3625..3948
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    3642..3698
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3699..3716)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3699..3715)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3708..3730)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3924..3943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    4079..4103
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4412..5041
                     /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 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
       61 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct gccggccctg
      121 caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa accatccggc
      181 ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa ggccgcgcag
      241 gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc
      301 gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgtc gtcgattagg
      361 aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta tgacgtgggc
      421 acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga
      481 cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt ttccgcaggg
      541 ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt ttcccatcta
      601 accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg cgtgttccgt
      661 ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa gcagaaagac
      721 gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca gcgtacgaag
      781 aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat tagccgctac
      841 aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct agctgattgg
      901 atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac
      961 tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc
     1021 aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag cgccggagag
     1081 ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct gccggagtac
     1141 gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg
     1201 atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg gcaaattgcc
     1261 ctagcagggg aaaaaggtcg aaaaagcttc tttcctgtgg atagcacgta cattgggaac
     1321 ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc gtacattggg
     1381 aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt ttccgcctaa
     1441 aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata actgtctggc
     1501 cagcgcacag ccgaacagct gcaaaaagcg cccacatcaa ggctccgagt gcgcggaacc
     1561 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1621 tgataaatgc ttcaataata ttgaaaaagg aagagtatgg ctaaaatgag aatatcaccg
     1681 gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct
     1741 gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc
     1801 cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa
     1861 ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg
     1921 ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct
     1981 gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg
     2041 gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat
     2101 aacgatctgg ccgatgtgga ttgcgaaaac tgggaagagg acactccatt taaagatccg
     2161 cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac
     2221 ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat
     2281 cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgctc
     2341 agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag
     2401 cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta gctgtcagac
     2461 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2521 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2581 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2641 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2701 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2761 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2821 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2881 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2941 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3001 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3061 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3121 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3181 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3241 ctgctcggat ctgttggacc ggacagtagt catggttgat gggctgcctg tatcgagtgg
     3301 tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca ggatatattg
     3361 tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt ttttaatgta
     3421 ctggggttga acactctgtg gaagacaatg cctgagaccg cagctggcac gacaggtttc
     3481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     3541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     3601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg
     3661 tcgactctag aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt
     3721 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3781 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3841 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3901 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3961 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4021 tacagacaag ctgtgaggtc tcagggacac gaagtgatcc gtttaaacta tcagtgtttg
     4081 acaggatata ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat
     4141 ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg
     4201 gttcccctcg ggagtcagcc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca
     4261 agctggcggc ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa
     4321 ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat
     4381 gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa
     4441 aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg
     4501 ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt
     4561 gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt
     4621 gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt
     4681 ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta
     4741 cgacatatgg gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga
     4801 tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg
     4861 tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca
     4921 gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga
     4981 gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg
     5041 agttaatgag gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg
     5101 agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag
     5161 cgggtcaact ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc
     5221 caaggcaaga ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg
     5281 agcaaatgaa taaatgagta gatgaatttt agc
//
