LOCUS       sequence-178457-        3606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap destination vector for building combined sgRNA expression + 
            repair template vectors, using the F+E sgRNA scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3606)
  AUTHORS   Dickinson D, Slabodnick M, Chen A, Goldstein B
  TITLE     SapTrap assembly of repair templates for Cas9-triggered homologous 
            recombination with a self-excising cassette.
  JOURNAL   MicroPubl Biol. 2018 May 1;2018:10.17912/W2KT0N. doi: 
            10.17912/W2KT0N.
  PUBMED    32550377
REFERENCE   2  (bases 1 to 3606)
  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: "MicroPubl 
            Biol."; date: "2018-05-1"; volume: "2018:10.17912/W2KT0N. doi"; 
            pages: " 10.17912/W2KT0N"
COMMENT     Sequence Label: pDD379
FEATURES             Location/Qualifiers
     source          1..3606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(433..782)
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     protein_bind    886..907
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(922..927,928..945,946..952)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    960..976
                     /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     965..987
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     984..1000
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     984..1000
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             996..1367
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAANSIGEEPMDPLVIWNSAGL
                     LFGNQAYRYRRPRGGARYPIRPIVSRITRAHWPSFYNVVTGKTLALPNLIALQHIPLSP
                     AGVIAKRPAPIALPNSCAA"
     primer_bind     1019..1039
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1021..1039
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1152..1168)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1153..1169)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1193..1212)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1194..1212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1222..1239)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1222..1238)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1231..1253)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1380..1835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1467..1486)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1677..1698
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1861..1965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1966..2034,2035..2826)
                     /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     complement(2184..2203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2997..3585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3486..3505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
       61 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
      121 acgaccgagc gcagcgagtc agtgagcgag gaagcgggtg tgaaataccg cacagatgcg
      181 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
      241 ggcgatcggt gcgggcctct tcgctattac gccagaaatt tcacaaaaaa aagcaccgac
      301 tcggtgccac tttttcaagt tgataacgga ctagccttat ttaaacttgc tatgctgttt
      361 ccagcatagc tcttaaaccg aagagcccat ggatccacta gtcatatgga attctgcagg
      421 cctgctcttc gcaagacatc tcgcaatagg aggtgtttat ataggggcaa cacaagaggc
      481 ggcagacatc tcgtgagttt gacagagtag tataaagagt gtgtgtgaga gggtgtaggc
      541 atttcataac atattttttg tggaataatt ttggaaaaaa ggtgtttttt taataatagc
      601 ttttataaga gtttctcaaa aattgcatgc tttttttgaa aaggttgaca aattatagta
      661 gtactaaagt tgctattttc aggtctcagc agaaaattca caactaaaaa taaaatttgc
      721 aaaacagagt gtgcgccttt aaggtgtaca gttttttatt cctttattgc tagttttttt
      781 tggcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      841 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      901 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      961 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
     1021 aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg gccaattcga
     1081 ttggcgaaga gcccatggat ccactagtca tatggaattc tgcaggcctg ctcttcggta
     1141 atcaagctta tcgataccgt cgacctcgag ggggggcccg gtacccaatt cgccctatag
     1201 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1261 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1321 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     1381 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1441 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1501 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1561 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1621 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1681 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1741 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1801 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     1861 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1921 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1981 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2041 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2101 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2161 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2221 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2281 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2341 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2401 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2461 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2521 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2581 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2641 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2701 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2761 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2821 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2881 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2941 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3001 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3061 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3121 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3181 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3241 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3301 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3361 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3421 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3481 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3541 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3601 gccttt
//
