LOCUS       sequence-136626-        2484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Simple cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2484)
  AUTHORS   Schwartz ML, Jorgensen EM
  TITLE     SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification
            in Caenorhabditis elegans.
  JOURNAL   Genetics. 2016 Feb 2. pii: genetics.115.184275.
  PUBMED    26837755
REFERENCE   2  (bases 1 to 2484)
  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: "Genetics. 
            2016 Feb 2. pii: genetics.115.184275."
COMMENT     Sequence Label: pMLS280
FEATURES             Location/Qualifiers
     source          1..2484
                     /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"
     primer_bind     563..582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        563..581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    590..697
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     606..622
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     607..623
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(657..673)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(657..673)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(710..730)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(710..728)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(749..765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(749..765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(867..886)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             878..1687
                     /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(945..964)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1834..2422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2323..2342
                     /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 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      601 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggggatc
      661 cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
      721 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctggcagc tctggcccgt
      781 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
      841 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
      901 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
      961 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1021 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1081 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1141 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1201 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1261 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1321 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1381 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     1441 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1501 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1561 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1621 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1681 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     1741 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     1801 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1861 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1921 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1981 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2041 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2101 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2161 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2221 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2281 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2341 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2401 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2461 ttgctggcct tttgctcaca tgtt
//
