LOCUS       sequence-490094-        4337 bp    DNA     circular SYN 21-OCT-2025
DEFINITION  Fragmid fragment: (Promoter) CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4337)
  TITLE     Fragmid
REFERENCE   2  (bases 1 to 4337)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AA662
FEATURES             Location/Qualifiers
     source          1..4337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     enhancer        448..827
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        829..1106
                     /label=chicken beta-actin promoter
     intron          1107..2123
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2047..2069
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2131..2150
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2247..2263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2247..2263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2260..2282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2271..2287
                     /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(2295..2301,2302..2319,2320..2325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2340..2361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2478..2495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2649..3237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2729..2748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3415..4224)
                     /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     4138..4157
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4216..4235
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg cagacggtca
       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 cacgctcgac attgattatt gactagttat taatagtaat
      481 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      541 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      601 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      661 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      721 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      781 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
      841 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
      901 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcccccg
      961 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
     1021 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
     1081 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
     1141 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
     1201 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
     1261 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
     1321 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct
     1381 ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca
     1441 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg
     1501 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt
     1561 cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     1621 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     1681 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
     1741 ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc
     1801 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc
     1861 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg
     1921 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1981 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
     2041 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
     2101 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     2161 attttggcaa agaattgagg agcgtcttca tcggatcccg ggcccgtcga ctgcagaggc
     2221 ctgcatgcaa gcttggtgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2281 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2341 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2401 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2461 gggcgctgtt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2521 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2581 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2641 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2701 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2761 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2821 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2881 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2941 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3001 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3061 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3361 agttttaaat caagcccaat ctgaataatg ttacaaccaa ttaaccaatt ctgattagaa
     3421 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     3481 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     3541 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     3601 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     3661 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     3721 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     3781 agccagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     3841 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     3901 taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg
     3961 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     4021 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     4081 tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc
     4141 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
     4201 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     4261 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     4321 cacgggccag agctgca
//
