LOCUS       sequence-8548-ex        3144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3144)
  AUTHORS   Merritt C, Seydoux G
  TITLE     Transgenic solutions for the germline.
  JOURNAL   WormBook. 2010 Feb 8:1-21. doi: 10.1895/wormbook.1.148.1.
  PUBMED    20169625
REFERENCE   2  (bases 1 to 3144)
  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"; journalName: "WormBook. 
            2010 Feb 8:1-21. doi: 10.1895/wormbook.1.148.1."
COMMENT     Sequence Label: pCM1.57
FEATURES             Location/Qualifiers
     source          1..3144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1190..1313
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1386..1404)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1409..1425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1409..1425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1527..1546)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1538..2347
                     /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     complement(1605..1624)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2494..3082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2983..3002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgct gcaggtcgac tctagaggat caccaaaaac
      721 ggaacgttga gctggacgga aatagtggta aagtgacatg attatagttt gaagatttct
      781 aatttcacaa ttagagcaaa tgttgttcgg tatttatttt caacggtatt tatactattt
      841 tccacctttt tctagaacat tcgagctgct tgttgcaaaa ggagggcgac tcacattcgg
      901 tacatggaaa agtagtgtac acaataaaga gacccagata cattttccgt ctgcgtctct
      961 ttgcacccac cgggagtatt ttcaaacgaa tgcatctagg accttctaga acattctgta
     1021 aggctgcaga atgcgggtat ataaggaaag cgggctcaga ggaagccaac acgctttgtt
     1081 ctagtgcatc taaaaaactt cgaaaatcct catcgggatc ctatcgattc gcggccgctg
     1141 tacacccggg tgccctatag tgagtcgtat tggtaccggt agaaaaaatc aagtttgtac
     1201 aaaaaagttg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     1261 cataaaaaac agactacata atactgtaaa acacaacata tgcagtcact atgaatcaac
     1321 tacttagatg gtattagtga cctgtagaat tcgagctcta gagctgcagg gcggccgcga
     1381 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1441 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1501 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1561 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1621 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1681 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1741 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1801 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1861 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1921 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1981 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2041 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2101 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2161 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2221 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2281 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2341 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2401 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2641 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2881 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3121 ttgctggcct tttgctcaca tgtt
//
