LOCUS       sequence-490102-        2642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Selection) for custom ORF cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2642)
  TITLE     Fragmid
REFERENCE   2  (bases 1 to 2642)
  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: AA692
FEATURES             Location/Qualifiers
     source          1..2642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(146..556)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAGLCSRRARDPMKTCFRILRRASSSSPVAPTSSSRC
                     IREVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNG
                     EWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(552..568)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(552..568)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(565..587)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    576..592
                     /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(600..606,607..624,625..630))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    645..666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(783..800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(954..1542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1034..1053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1720..2529)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2443..2462
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2521..2540
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacgta ggagccaccg gagacgatga gctagcccgt ctcaggatcc
      481 ggaagcacgt cttcatcgga tcccgggccc gtcgactgca gaggcctgca tgcaagcttg
      541 gtgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      601 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      661 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      721 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      781 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      841 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      901 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      961 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1021 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1081 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1141 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1201 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1261 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1321 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1381 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1441 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1501 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1561 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1621 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaagc
     1681 ccaatctgaa taatgttaca accaattaac caattctgat tagaaaaact catcgagcat
     1741 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     1801 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     1861 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     1921 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     1981 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     2041 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     2101 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     2161 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     2221 tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     2281 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     2341 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     2401 cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     2461 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat
     2521 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2581 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg gccagagctg
     2641 ca
//
