LOCUS       sequence-314764-        2998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  places LacZalpha unit between Esp3I sites for better cutting of 
            plasmid and ability to distinguish recombinants as white colonies 
            when plated on Xgal plates.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2998)
  TITLE     Modified donor plasmids to enhance Golden Gate assembly of 
            Drosophila two stage Crispr/Cas9 cassette exchange
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2998)
  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            "Unpublished"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: pXZ13lacZalpha
FEATURES             Location/Qualifiers
     source          1..2998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    566..587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(602..607,608..625,626..632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    640..656
                     /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)."
     primer_bind     645..667
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     664..680
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     664..680
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             676..924
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRSNIQ"
     misc_feature    690..746
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(750..767)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(750..766)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(759..781)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1080..1097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1251..1839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1331..1350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2010..2801,2802..2870))
                     /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"
     primer_bind     2633..2652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2871..2975)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa atgagctgat ttaacaaaaa
      421 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
      481 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ggtacggact
      541 gagacgtgag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
      601 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
      661 acacaggaaa cagctatgac catgattacg aattcgagct cggtacccgg ggatcctcta
      721 gagtcgacct gcaggcatgc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
      781 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
      841 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
      901 gaatggcgtt ccaacatcca ataaatcata caggcaaggc aaagaattag caaaattaag
      961 caataaagcc tcagagcata aagctaaatc ggttgtacca cgtctcaatg caggctgcat
     1021 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1081 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1141 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1201 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1261 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1321 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1381 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1441 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1501 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1561 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1621 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1681 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1741 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1801 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1861 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1921 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1981 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2041 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2101 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2161 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2221 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2281 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2341 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2401 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2461 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2521 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2581 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2641 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2701 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2761 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2821 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2881 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2941 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
