LOCUS       sequence-469973-        3382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR cloning, gRNA expression, and detection of undesired vector 
            insertion during HDR for Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3382)
  TITLE     McCabe Lab Plasmids 2025
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3382)
  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            "Unpublished"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: pVID
FEATURES             Location/Qualifiers
     source          1..3382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      220..229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1043..1164
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1080..1099)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1134..1153
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1165..1585
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     rep_origin      complement(1627..2215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1707..1726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2386..3177,3178..3246))
                     /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     3009..3028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3247..3351)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatctaatt caattagaga ctaattcaat tagagctaat tcaattagga tccaagctta
       61 tcgatttcga accctcgacc gccggagtat aaatagaggc gcttcgtcta cggagcgaca
      121 attcaattca aacaagcaaa gtgaacacgt cgctaagcga aagctaagca aataaacaag
      181 cgcagctgaa caagctaaac aatcggctcg aagccggtcg ccaccatggc ccaaagcaag
      241 cacggcctga ctaaagaaat gacaatgaag tacaggatgg aaggttgcgt tgatggtcac
      301 aaatttgtca taactggaga aggcataggt tatccgttca aaggcaagca ggcgatcaac
      361 ctgtgtgtag tggaaggagg ccctctccca tttgcggaag acatcttgag tgctgccttc
      421 atgtatggta accgggtttt tactgaatac cctcaggata tagccgatta tttcaagaac
      481 tcctgtcccg ccggctatac ttgggaccgg tcgttcttgt ttgaggatgg tgctgtctgt
      541 atctgcaacg cggacattac agtatcggtg gaggaaaact gtatgtacca tgaaagcaaa
      601 ttttacggcg taaattttcc agccgacggc cccgtcatga agaagatgac agacaactgg
      661 gagcctagtt gcgagaaaat aattcctgtt cccaagcagg gtatcctcaa gggcgacgtt
      721 agtatgtact tgctcctgaa agacggtggc cggctgcgct gccaatttga tactgtttat
      781 aaggccaaga gtgtgcctag gaagatgccc gactggcatt ttatacaaca taagttgacc
      841 cgcgaggacc ggagtgacgc caaaaaccag aaatggcatc tcacagaaca cgccattgca
      901 agcggaagtg cgttgccctg agcggccgcg actctagatc ataatcagcc ataccacatt
      961 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1021 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1081 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1141 gtccaaactc atcaatgtat cttatttttt gctcacctgt gattgctcct actcaaatac
     1201 aaaaacatca aattttctgt caataaagca tatttattta tatttatttt acaggaaaga
     1261 attcctttta aagtgtattt taacctataa tgaaaaacga ttaaaaaaaa tacataaaat
     1321 aattcgaaaa tttttgaata gcccaggttg ataaaaattc atttcatacg ttttataact
     1381 tatgccccta agtatttttt gaccatagtg tttcaattct acattaattt tacagagtag
     1441 aatgaaacgc cacctactca gccaagaggc gaaaaggtta gctcgccaag cagagagggc
     1501 gccagtgctc actacttttt ataattctca acttcttttt ccagactcag ttcgtatata
     1561 tagacctatt ttcaatttaa cgtcgaggcc tgcagcccgg gccggccctc gagatttaaa
     1621 tgcatttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2041 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2101 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2161 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2221 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2281 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2341 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2461 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2521 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2581 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2821 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2881 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2941 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3001 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3061 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3241 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3301 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3361 ccgaaaagtg ccacctgacg tc
//
