LOCUS       sequence-343537-        2844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 Acceptor for creating N-tag modules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2844)
  TITLE     Banfield lab plasmids
REFERENCE   2  (bases 1 to 2844)
  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: pICSL01002
FEATURES             Location/Qualifiers
     source          1..2844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(216..1007)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1229..1333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1401..1419
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    1520..1541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1556..1561,1562..1579,1580..1586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1594..1610
                     /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     1599..1621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1618..1634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1618..1634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1630..1953
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    1647..1703
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1704..1721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1704..1720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1713..1735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1929..1948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2130..2147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2301..2844,1..45))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2381..2400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
       61 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      121 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      181 atctaaagta tatatgagta aacttggtct gacagttatt tgccgactac cttggtgatc
      241 tcgcctttta cgtaatggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
      301 tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc
      361 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
      421 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc
      481 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
      541 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
      601 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
      661 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
      721 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
      781 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
      841 acggtcaccg taaccagcaa atcaatatcg ctgtgtggct tcaggccgcc atccactgcg
      901 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
      961 acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatgat gtttaacttt
     1021 gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
     1081 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac cccaaaaaaa
     1141 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     1201 aggttctgga ccagttgcgt gagcgcatac tcttcctttt tcaatattat tgaagcattt
     1261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     1321 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     1381 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcca ctgaagagcc
     1441 actgtggtct caccatttgt cttcgcagct ggcacgacag gtttcccgac tggaaagcgg
     1501 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     1561 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     1621 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat
     1681 ccccgggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1861 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     1921 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     1981 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2041 acgaagacaa aatgtgagac cacgagtgat taatgaatcg gccaacgcgc ggggagaggc
     2101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2821 accaccgctg gtagcggtgg tttt
//
