LOCUS       sequence-234019-        3911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 acceptor vector for promoters with lacZ compatible with 
            blue/white screening, low copy number, pSC101 ori. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3911)
  AUTHORS   Vasudevan R, Gale GAR, Schiavon AA, Puzorjov A, Malin J, Gillespie 
            MD, Vavitsas K, Zulkower V, Wang B, Howe CJ, Lea-Smith DJ, McCormick
            AJ
  TITLE     CyanoGate: A Modular Cloning Suite for Engineering Cyanobacteria 
            Based on the Plant MoClo Syntax.
  JOURNAL   Plant Physiol. 2019 May;180(1):39-55. doi: 10.1104/pp.18.01401. Epub
            2019 Feb 28.
  PUBMED    30819783
REFERENCE   2  (bases 1 to 3911)
  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"; doi: 
            "10.1104/pp.18.01401"; journalName: "Plant Physiol"; date: 
            "2019-05"; volume: "180"; issue: "1"; pages: "39-55"
COMMENT     Sequence Label: pCA0.002
FEATURES             Location/Qualifiers
     source          1..3911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..43
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(142..957)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     865..884
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     949..968
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1315..2265)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2313..2535
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3027..3121)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(3277..3306)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     protein_bind    3381..3402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3417..3422,3423..3440,3441..3447)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3455..3471
                     /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     3460..3482
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3479..3495
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3479..3495
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3491..3814
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    3508..3564
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3565..3582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3565..3581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3574..3596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3790..3809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aatgtgagac cagtcaaaag cctccgaccg gaggcttttg actaaaactt cccttggggt
       61 tatcattggg gctcactcaa aggcggtaat cagataaaaa aaatccttag ctttcgctaa
      121 ggatgatttc tgctagtatt attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
      181 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
      241 aaactcaccg aggcagttcc aaagaatggc aaggtcctgg taacggtctg cgattccgac
      301 ccgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
      361 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aagagcttgt gcatttcttt
      421 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
      481 accgttattc atgcgtgatt gcgcctgagc aagacgaaat acacgatcgc tgttaaaagg
      541 acaattacaa acaggaatcg aatgtaaccg gcgcaggaac acggccagcg catcaacaat
      601 attttcacct gaatcaggat attcttctaa tacctggaag gctgttttcc caggaatcgc
      661 ggtggtgagt aaccacgcat catcaggagt acggataaaa tgcttgatgg tcgggagagg
      721 cataaactcc gtcagccagt tgagacggac catctcatct gtaacatcat tggcaacgct
      781 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat
      841 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcgtc
      901 catgttggag tttaagcgcg gacgggagca agacgtttcc cgttgaatat ggctcataac
      961 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     1021 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     1081 aacttttgct gagttgaagg atcagctcta gtagttacat tgtcgatctg ttcatggtga
     1141 acagctttga atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg
     1201 ttttcatctg tgcatatgga cagttttccc tttgatatgt aacggtgaac agttgttcta
     1261 cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat
     1321 ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga
     1381 gaacgaacca ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact
     1441 ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag
     1501 gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt
     1561 ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc
     1621 agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt
     1681 acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag
     1741 cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt
     1801 ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga
     1861 cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat
     1921 ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg
     1981 gaaaatctca aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc
     2041 tctggttgct ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc
     2101 gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg
     2161 gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg
     2221 actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc
     2281 taggtgattt taatcactat accaattgag atgggctagt caatgataat tacatgtcct
     2341 tttcctttga gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa
     2401 ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca
     2461 agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag
     2521 ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac
     2581 gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc
     2641 aagctcgggc aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct
     2701 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat
     2761 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc
     2821 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt
     2881 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt
     2941 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta
     3001 cccgtcttac tgtccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     3061 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     3121 ccaagcgagc tcgtaaactt ggtctgacag ctctagctcc ggcaaaaaaa cgggcaaggt
     3181 gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgttt gccacctgac
     3241 gtctaagaaa aggaatattc agcaatttgc ccgtgccgaa gaaaggccca cccgtgaagg
     3301 tgagccggtc tcaggagttg tcttcgcagc tggcacgaca ggtttcccga ctggaaagcg
     3361 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ctaggcttta
     3421 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     3481 ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga
     3541 tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     3601 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     3661 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3721 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     3781 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3841 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3901 gacgaagaca a
//
