LOCUS       sequence-274775-        6370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used in the editing of Drosophila kkv.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6370)
  AUTHORS   Adler PN
  TITLE     The localization of chitin synthase mediates the patterned 
            deposition of chitin in developing Drosophila bristles
  JOURNAL   bioRxiv 2020
REFERENCE   2  (bases 1 to 6370)
  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 
            2020"
COMMENT     Sequence Label: pCFD3-dUG-D01
FEATURES             Location/Qualifiers
     source          1..6370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    34..419
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     promoter        433..853
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     misc_RNA        874..949
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1260..1280)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1260..1278)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1299..1315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1299..1315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1312..1334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1323..1339
                     /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)."
     promoter        complement(join(1347..1353,1354..1371,1372..1377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1392..1413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1530..1547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1701..2289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1781..1800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2460..3251,3252..3320))
                     /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     3083..3102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3321..3425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3452..3908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3539..3558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3749..3770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4034..4056
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4048..4065
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4049..4065
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4075..4094
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4075..4093
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(4285..4402,4479..4572,4664..5270,5325..5458,
                     5519..5679,5753..5778))
                     /codon_start=1
                     /product="tryptophan oxygenase"
                     /label=vermilion
                     /note="selectable marker, required to synthesize the brown 
                     eye pigment in Drosophila"
                     /translation="MSCPYAGNGNDHDDSAVPLTTEVGKIYGEYLMLDKLLDAQCMLSE
                     EDKRPVHDEHLFIITHQAYELWFKQIIFEFDSIRDMLDAEVIDETKTLEIVKRLNRVVL
                     ILKLLVDQVPILETMTPLDFMDFRKYLAPASGFQSLQFRLIENKLGVLTEQRVRYNQKY
                     SDVFSDEEARNSIRNSEKDPSLLELVQRWLERTPGLEESGFNFWAKFQESVDRFLEAQV
                     QSAMEEPVEKAKNYRLMDIEKRREVYRSIFDPAVHDALVRRGDRRFSHRALQGAIMITF
                     YRDEPRFSQPHQLLTLLMDIDSLITKWRYNHVIMVQRMIGSQQLGTGGSSGYQYLRSTL
                     SDRYKVFLDLFNLSTFLIPREAIPPLDETIRKKLINKSV"
     protein_bind    6093..6162
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 cactagaagg cctaattcgg tacactagtt ggccacgtaa taagtgtgcg ttgaatttat
       61 tcgcaaaaac attgcatatt ttcggcaaag taaaattttg ttgcatacct tatcaaaaaa
      121 taagtgctgc atacttttta gagaaaccaa ataatttttt attgcatacc cgtttttaat
      181 aaaatacatt gcataccctc ttttaataaa aaatattgca tactttgacg aaacaaattt
      241 tcgttgcata cccaataaaa gattattata ttgcataccc gtttttaata aaatacattg
      301 cataccctct tttaataaag aatattgcat acgttgacga aacaaatttt cgttgcatac
      361 ccaataaaag attattatat tgcatacctt ttcttgccat accatttagc cgatcaattg
      421 agatctgaat tcttttttgc tcacctgtga ttgctcctac tcaaatacaa aaacatcaaa
      481 ttttctgtca ataaagcata tttatttata tttattttac aggaaagaat tccttttaaa
      541 gtgtatttta acctataatg aaaaacgatt aaaaaaaata cataaaataa ttcgaaaatt
      601 tttgaatagc ccaggttgat aaaaattcat ttcatacgtt ttataactta tgcccctaag
      661 tattttttga ccatagtgtt tcaattctac attaatttta cagagtagaa tgaaacgcca
      721 cctactcagc caagaggcga aaaggttagc tcgccaagca gagagggcgc cagtgctcac
      781 tactttttat aattctcaac ttctttttcc agactcagtt cgtatatata gacctatttt
      841 caatttaacg tcgtttctca ctctcatcaa actgttttag agctagaaat agcaagttaa
      901 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgccta
      961 cctggagcct gagagttgtt caataaaata aaaatgtttc gtttttttgc tttcgccagt
     1021 atttattatt tttcatcaat atgtattcaa tttggtatgt atttagtaat tgtaatatat
     1081 agacaatggt tttccgttga cgtacataca tctgacgtgt gtttatttag acataatagt
     1141 tatgttttca catcttttta atgttcgctt aatgcgtatg cattctagac aattgtgctc
     1201 ggcaacagta tatttgtggt gtgccaacca acaacctgca ggagctccag cttttgttcc
     1261 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     1321 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     1381 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     1441 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1501 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1561 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1621 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2101 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2401 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2461 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2521 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2581 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2641 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2701 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2761 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2821 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2881 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2941 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3001 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3061 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3121 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3181 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3241 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3301 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3421 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     3481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3661 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3781 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3841 ttaaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3901 tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     3961 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     4021 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     4081 gactcactat agggcgaatt gggtacaagc ttggatttat tttgttatgt tatatgtatt
     4141 atatgtcaga cataaagaaa aggaacacat caaatgtgat aacaaagact aaacaagtaa
     4201 ttttattaca ccaaaacgac aaaacagtag gcagaacaaa caacgcatag ccaaacattg
     4261 acgaattgga taccctgccg attgtcagac acttttgttg atcagtttct tgcgaatggt
     4321 ctcgtccagc ggtggaatcg cctcgcgggg aatcagaaaa gtggacagat tgaacagatc
     4381 cagaaacacc ttgtaccgat cactgaaacc aaaaaaaaac aaagggagaa cagtttgagt
     4441 tcattgatcc ccgatataat cacatctgcg atgatcacct gagagtggag cgcagatatt
     4501 gatatccaga cgagccacca gtgcccaact gttgggatcc aatcatgcgt tgcaccatga
     4561 tcacgtgatt gtctgcggcg ggaatagaaa gtatttggtt aggaaaacca gtcttaaaca
     4621 taagatatat ttataaaaga gtatcaaaga atgcaatact tacatctcca cttggttatt
     4681 aacgagtcga tgtccatgag cagggtgagc aactggtgtg gttggctgaa cctgggttca
     4741 tccctataga aggtgatcat gatggctccc tgaagggcac gatggctaaa ccggcgatcc
     4801 ccacgacgca ccagtgcatc gtgcactgcc ggatcaaaga tggagcgata cacctcgcgt
     4861 cgcttctcaa tgtccatgag gcggtagttt ttcgccttct ccacgggctc ctccatggcg
     4921 ctctgtacct gcgcctccag gaatcgatcg acgctctcct gaaacttggc ccagaagttg
     4981 aagccactct cctccagtcc gggcgtcctc tccagccatc gctgcactag ctccagtagc
     5041 gagggatctt tctccgagtt gcgaatcgag ttccgcgcct cctcgtcgct aaagacatcc
     5101 gagtacttct ggttgtatct cacccgctgc tctgtcagaa ctcccagctt gttctcgatc
     5161 aaacggaact gcagcgactg aaaaccagat gcgggtgcca ggtacttgcg gaagtccatg
     5221 aagtctagcg gggtcatggt ctccagaatg ggcacttggt ccaccaggag ctgtacaaag
     5281 gaagttataa acggattttg gtaagagatt cagaaagcac tcacttttag aatcagaacc
     5341 actcggttca gtcgcttgac aatctccagc gtcttggttt catcgatgac ctctgcatcc
     5401 aacatgtctc gtatggagtc gaactcaaag atgatctgct tgaaccaaag ctcgtaggct
     5461 gtggcgaagg tacttaaatg ccattgagtg ttgtcatcaa agttgtaaac ctactcaccc
     5521 tggtgcgtga tgatgaacag atgctcatcg tgcacgggtc gcttgtcctc ctcggacagc
     5581 atacactggg catccagcag tttgtccagc atcagatact ctccatagat tttgcccact
     5641 tccgtggtta atggcaccgc cgaatcatcg tgatcgtttc tgtatgggtt tgaattgaat
     5701 cgcagaactg aagatcgatt ggcattcctg gacagcacgt gctggtgctc acccgtttcc
     5761 tgcataggga cagctcatgg tgcacagctc agatcagatc gtgactcctc gagcggcgga
     5821 tgctggcgaa ctgatctccg ccagcggacc ggagatgaga ccccagcgaa ccgataacag
     5881 agcgagagag ctccagttcc gactgattgc acagtcggtg atctgggcga tgggcactgc
     5941 cagataggct gggaattatc aatcacttga ggtgaaagtg cggcgcacac aaatccaagc
     6001 ttgatatcat cgatctcgag gctgcatcca acgcgttggg agctctccgg atcaattcgg
     6061 cttcaggtac cgtcgacgat gtaggtcacg gtctcgaagc cgcggtgcgg gtgccagggc
     6121 gtgcccttgg gctccccggg cgcgtactcc acctcaccca tctggtccat catgatgaac
     6181 gggtcgaggt ggcggtagtt gatcccggcg aacgcgcggc gcaccgggaa gccctcgccc
     6241 tcgaaaccgc tgggcgcggt ggtcacggtg agcacgggac gtgcgacggc gtcggctggt
     6301 gcggatacgc ggggcagcgt cagcgggttc tcgacggtca cggcgggcat gtcgacaagc
     6361 cgaattgatc
//
