LOCUS       sequence-152786-        5869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating dsDNA donors for HDR. Empty backbone 
            containing a StuI restriction site and GMR-white for identifying 
            backbone incorporation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5869)
  TITLE     Kate O'Connor-Giles Lab Donor Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5869)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pHD-w+
FEATURES             Location/Qualifiers
     source          1..5869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     618..637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(856..947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     989..1008
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1014..1033
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1824..1841)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1824..1840)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1939..1956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2110..2698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2190..2209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5864..5869,1..786,787..855))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
       61 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagcgc
      121 tgcgatgata ccgcgagaac cacgctcacc ggctccggat ttatcagcaa taaaccagcc
      181 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      241 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      301 tgccatcgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      361 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      421 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      481 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
      541 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
      601 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
      661 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
      721 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
      781 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
      841 atgttgaata ctcatattct tcctttttca atattattga agcatttatc agggttattg
      901 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt
      961 tacaaccaat taaccaattc tgaacattat cgcgagccca tttatacctg aatatggctc
     1021 ataacacccc ttgtttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1081 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gactccccat gcgagagtag
     1141 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgcccg
     1201 ggctaataag gggggctgga tcgcttcgtg ttccccatcg gtgatgtcgt ataggaagca
     1261 gtatacgaga cctataggag acgtatatgg tcttcttttc ccgaggcctc ggccccgaag
     1321 acactatagg tctcctatac gtctcttata cgacatcacc gatggggaac ccagacgctg
     1381 agtacgtatt ctaaatgcat aataaatact gataacatct tatagtttgt attatatttt
     1441 gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt tattattttc
     1501 gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac aaaaaatcat
     1561 aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca acgtatctta
     1621 tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata tatcaagcaa
     1681 agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc ccccataaaa
     1741 aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg ttatcagaac
     1801 cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa gagtttgtag
     1861 aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct
     1921 actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2521 gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca
     2761 cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcgtacga ctagcacctg
     2821 actgtcgagc ccagtggaaa cccttgaaat gcctttaagt cgagcccagt ggaaaccctt
     2881 gaaatgcctt taagtcgagc ccagtggaaa cccttgaaat gcctttaagt cgagcccagt
     2941 ggaaaccctt gaaatgcctt taagtcgagc ccagtggaaa cccttgaaat gcctttaagt
     3001 cgagtagcag tcgaggtagg tacgtcatgc cccgttattc tctattcgtt ttgtgactct
     3061 ccctctctgt actattgctc tctcactctg tcgcacagta aacggcactc tattctcgtt
     3121 gcttcgagag agcgcgcctc gaatgttcgc gaaaagagcg ccggagtata aatagaggcg
     3181 cttcgtcgac ggagcgtcaa ttcaattcaa acaagcaaag tgaacacatc gcgaagcaag
     3241 ctgagcaaac aaacaagcgc agctgaacaa gctaaacaat ctgcaataaa gtgcaagtta
     3301 aagtgaatca attaaaagta accaacaacc aagtaattaa actaaaaact gcaactactg
     3361 aaatcaacca agaagtcatt attgaagaca agaagagaac tctgaatagg tcgatagcgt
     3421 caattcggca cgaggtttcg tgacgaagct ccaagcggtt tacgccatca attaaacaca
     3481 aagtgctgtg ccaaaactcc tctcgcttct tatttttgtt tgttttttga gtgattgggt
     3541 gggtgattgg ttttgggtgg gtaagcaggg gaaagtgtga aaaatcccgg caatgggcca
     3601 agaggatcag gagctattaa ttcgcggagg cagcaaacac ccatctgccg agcatctgaa
     3661 caatggtgac agcggagcgg cttcgcagag ctgcattaac cagggcttcg ggcaggccaa
     3721 aaactacggc acgctcctgc cacccagtcc gccggaggac tccggttcag ggagcggcca
     3781 actagccgag aacctcacct atgcctggca caatatggac atctttgggg cggtcaatca
     3841 gccgggctcc ggatggcggc agctggtcaa ccggacacgc ggactattct gcaacgagcg
     3901 acacataccg gcgcccagga aacatttgct caagaacgtt tgcggcgtgg cctatccggg
     3961 cgaacttttg gccgtgatgg gcagttccgg tgccggaaag acgaccctgc tgaatgccct
     4021 tgcctttcga tcgccgcagg gcatccaagt atcgccatcc gggatgcgac tgctcaatgg
     4081 ccaacctgtg gacgccaagg agatgcaggc caggtgcgcc tatgtccagc aggatgacct
     4141 ctttatcggc tccctaacgg ccagggaaca cctgattttc caagccatgg tgcggatgcc
     4201 acgacatctg acctatcggc agcgagtggc ccgcgtggat caggtgatcc aggagctttc
     4261 gctcagcaaa tgtcagcaca cgatcatcgg tgtgcccggc agggtgaaag gtctgtccgg
     4321 cggagaaagg aagcgtctgg cattcgcctc cgaggcacta accgatccgc cgcttctgat
     4381 ctgcgatgag cccacctccg gactggactc atttaccgcc cacagcgtcg tccaggtgct
     4441 gaagaagctg tcgcagaagg gcaagaccgt catcctgacc attcatcagc cgtcttccga
     4501 gctgtttgag ctctttgaca agatccttct gatggccgag ggcagggtag ctttcttggg
     4561 cactcccagc gaagccgtcg acttcttttc ctacgtgggt gcccagtgtc ctaccaacta
     4621 caatccggcg gacttttacg tacaggtgtt ggccgttgtg cccggacggg agatcgagtc
     4681 ccgtgatcgg atcgccaaga tatgcgacaa ttttgccatt agcaaagtag cccgggatat
     4741 ggagcagttg ttggccacca aaaatctgga gaagccactg gagcagccgg agaatgggta
     4801 cacctacaag gccacctggt tcatgcagtt ccgggcggtc ctgtggcgat cctggctgtc
     4861 ggtgctcaag gaaccactcc tcgtaaaagt gcgacttatt cagacaacga tggttgccat
     4921 cttgattggc ctcatctttt tgggccaaca actcacgcaa gtgggtgtga tgaatatcaa
     4981 cggagccatc ttcctcttcc tgaccaacat gacctttcaa aacgtctttg ccacgataaa
     5041 tgtgttcacc tcagagctgc cagtttttat gagggaggcc cgaagtcgac tttatcgctg
     5101 tgacacatac tttctgggca aaacgattgc cgaattgccg ctttttctca cagtgccact
     5161 ggtcttcacg gcgattgcct atccgatgat cggactgcgg gccggagtgc tgcacttctt
     5221 caactgcctg gcgctggtca ctctggtggc caatgtgtca acgtccttcg gatatctaat
     5281 atcctgcgcc agctcctcga cctcgatggc gctgtctgtg ggtccgccgg ttatcatacc
     5341 attcctgctc tttggcggct tcttcttgaa ctcgggctcg gtgccagtat acctcaaatg
     5401 gttgtcgtac ctctcatggt tccgttacgc caacgagggt ctgctgatta accaatgggc
     5461 ggacgtggag ccgggcgaaa ttagctgcac atcgtcgaac accacgtgcc ccagttcggg
     5521 caaggtcatc ctggagacgc ttaacttctc cgccgccgat ctgccgctgg actacgtggg
     5581 tctggccatt ctcatcgtga gcttccgggt gctcgcatat ctggctctaa gacttcgggc
     5641 ccgacgcaag gagtagccga catatatccg aaataactgc ttgttttttt tttttaccat
     5701 tattaccatc gtgtttactg tttattgccc cctcaaaaag ctaatgtaat tatatttgtg
     5761 ccaataaaaa caagatatga cctatagaat acaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5821 actcgacacc gggatccaaa cctagcagcg taatgctctg cttttacca
//
