LOCUS       sequence-397950-        2465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Even assembly end-linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2465)
  AUTHORS   Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z,
            Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N
  TITLE     A scalable framework for high-throughput identification of 
            functional origins of replication in non-model bacteria
  JOURNAL   bioRxiv 2023.05.19.541510
REFERENCE   2  (bases 1 to 2465)
  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: "bioRxiv 
            2023.05.19.541510"
COMMENT     Sequence Label: pGLEL_3 [epsilon-omega]
FEATURES             Location/Qualifiers
     source          1..2465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(357..373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(357..373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(370..392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(545..1133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(625..644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1269..2084)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1269..2084)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1992..2011
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2076..2095
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2205..2221
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2205..2221
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      2222..2268
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2382..2404
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2396..2413
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2397..2413
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gactagttgg aagtttgtct agatctcaga ttttgtcact agaggacgca cgctctattt
       61 ttatgatcca ttgatgtccc tgacgctgca aaatttgcaa ccaggcagtc ttcgcggtag
      121 gtcctagtgc aatggggctt tttttccata gtcctcgaga ggaggagacg tcagtccaga
      181 tatctttgat gtcgtgattg gaaggaccct tggccctcca cccttaggca gtgttggctc
      241 ttcacagctg acctcctgcc agcaatagta agacaacacg caaagtcggt tgaagagcgt
      301 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
      361 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
      421 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
      481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1141 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
     1201 agcgtaaatg ggtagggggc ttcaaatcgt ccgctctgcc agtgttacaa ccaattaaca
     1261 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     1321 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     1381 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     1441 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     1501 gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca
     1561 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     1621 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     1681 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     1741 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     1801 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     1861 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     1921 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     1981 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     2041 aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     2101 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     2161 taacatcaga gattttgaga cacaacgtgg ctttcccccg ccgctctaga actagtggat
     2221 ccaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     2281 gcattatacg agacgtccag gttgggatac ctgaaacaaa acccatcgta cggccaagga
     2341 agtctccaat aactgtgatc caccacaagc gccagggttt tcccagtcac gacgttgtaa
     2401 aacgacggcc agtcatgcat aatccgcacg catctggaat aaggaagtgc cattccgcct
     2461 gacct
//
