LOCUS       sequence-274780-        6370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to edit the endogenous Drosophila kkv gene to encode a 
            fluorescent Chitin Synthase..
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-DH1
FEATURES             Location/Qualifiers
     source          1..6370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    35..420
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     promoter        434..854
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     misc_RNA        875..950
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1261..1281)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1261..1279)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1300..1316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1300..1316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1313..1335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1324..1340
                     /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(1348..1354,1355..1372,1373..1378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1393..1414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1531..1548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1702..2290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1782..1801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2461..3252,3253..3321))
                     /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     3084..3103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3322..3426)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3453..3909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3540..3559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3750..3771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4035..4057
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4049..4066
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4050..4066
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4076..4095
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4076..4094
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(4286..4403,4480..4573,4665..5271,5326..5459,
                     5520..5680,5754..5779))
                     /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    6094..6163
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 ccactagaag gcctaattcg gtacactagt tggccacgta ataagtgtgc gttgaattta
       61 ttcgcaaaaa cattgcatat tttcggcaaa gtaaaatttt gttgcatacc ttatcaaaaa
      121 ataagtgctg catacttttt agagaaacca aataattttt tattgcatac ccgtttttaa
      181 taaaatacat tgcataccct cttttaataa aaaatattgc atactttgac gaaacaaatt
      241 ttcgttgcat acccaataaa agattattat attgcatacc cgtttttaat aaaatacatt
      301 gcataccctc ttttaataaa gaatattgca tacgttgacg aaacaaattt tcgttgcata
      361 cccaataaaa gattattata ttgcatacct tttcttgcca taccatttag ccgatcaatt
      421 gagatctgaa ttcttttttg ctcacctgtg attgctccta ctcaaataca aaaacatcaa
      481 attttctgtc aataaagcat atttatttat atttatttta caggaaagaa ttccttttaa
      541 agtgtatttt aacctataat gaaaaacgat taaaaaaaat acataaaata attcgaaaat
      601 ttttgaatag cccaggttga taaaaattca tttcatacgt tttataactt atgcccctaa
      661 gtattttttg accatagtgt ttcaattcta cattaatttt acagagtaga atgaaacgcc
      721 acctactcag ccaagaggcg aaaaggttag ctcgccaagc agagagggcg ccagtgctca
      781 ctacttttta taattctcaa cttctttttc cagactcagt tcgtatatat agacctattt
      841 tcaatttaac gtcgctggtg gtgcgggtgg ggatgtttta gagctagaaa tagcaagtta
      901 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgcct
      961 acctggagcc tgagagttgt tcaataaaat aaaaatgttt cgtttttttg ctttcgccag
     1021 tatttattat ttttcatcaa tatgtattca atttggtatg tatttagtaa ttgtaatata
     1081 tagacaatgg ttttccgttg acgtacatac atctgacgtg tgtttattta gacataatag
     1141 ttatgttttc acatcttttt aatgttcgct taatgcgtat gcattctaga caattgtgct
     1201 cggcaacagt atatttgtgg tgtgccaacc aacaacctgc aggagctcca gcttttgttc
     1261 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1501 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2101 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2281 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2341 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2401 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2461 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2521 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2581 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2641 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2701 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2761 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2821 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2881 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2941 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3001 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3061 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3121 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3181 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3241 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3301 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3421 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     3481 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3541 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3601 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3661 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3721 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3781 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3841 gttaaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3901 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3961 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     4021 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     4081 cgactcacta tagggcgaat tgggtacaag cttggattta ttttgttatg ttatatgtat
     4141 tatatgtcag acataaagaa aaggaacaca tcaaatgtga taacaaagac taaacaagta
     4201 attttattac accaaaacga caaaacagta ggcagaacaa acaacgcata gccaaacatt
     4261 gacgaattgg ataccctgcc gattgtcaga cacttttgtt gatcagtttc ttgcgaatgg
     4321 tctcgtccag cggtggaatc gcctcgcggg gaatcagaaa agtggacaga ttgaacagat
     4381 ccagaaacac cttgtaccga tcactgaaac caaaaaaaaa caaagggaga acagtttgag
     4441 ttcattgatc cccgatataa tcacatctgc gatgatcacc tgagagtgga gcgcagatat
     4501 tgatatccag acgagccacc agtgcccaac tgttgggatc caatcatgcg ttgcaccatg
     4561 atcacgtgat tgtctgcggc gggaatagaa agtatttggt taggaaaacc agtcttaaac
     4621 ataagatata tttataaaag agtatcaaag aatgcaatac ttacatctcc acttggttat
     4681 taacgagtcg atgtccatga gcagggtgag caactggtgt ggttggctga acctgggttc
     4741 atccctatag aaggtgatca tgatggctcc ctgaagggca cgatggctaa accggcgatc
     4801 cccacgacgc accagtgcat cgtgcactgc cggatcaaag atggagcgat acacctcgcg
     4861 tcgcttctca atgtccatga ggcggtagtt tttcgccttc tccacgggct cctccatggc
     4921 gctctgtacc tgcgcctcca ggaatcgatc gacgctctcc tgaaacttgg cccagaagtt
     4981 gaagccactc tcctccagtc cgggcgtcct ctccagccat cgctgcacta gctccagtag
     5041 cgagggatct ttctccgagt tgcgaatcga gttccgcgcc tcctcgtcgc taaagacatc
     5101 cgagtacttc tggttgtatc tcacccgctg ctctgtcaga actcccagct tgttctcgat
     5161 caaacggaac tgcagcgact gaaaaccaga tgcgggtgcc aggtacttgc ggaagtccat
     5221 gaagtctagc ggggtcatgg tctccagaat gggcacttgg tccaccagga gctgtacaaa
     5281 ggaagttata aacggatttt ggtaagagat tcagaaagca ctcactttta gaatcagaac
     5341 cactcggttc agtcgcttga caatctccag cgtcttggtt tcatcgatga cctctgcatc
     5401 caacatgtct cgtatggagt cgaactcaaa gatgatctgc ttgaaccaaa gctcgtaggc
     5461 tgtggcgaag gtacttaaat gccattgagt gttgtcatca aagttgtaaa cctactcacc
     5521 ctggtgcgtg atgatgaaca gatgctcatc gtgcacgggt cgcttgtcct cctcggacag
     5581 catacactgg gcatccagca gtttgtccag catcagatac tctccataga ttttgcccac
     5641 ttccgtggtt aatggcaccg ccgaatcatc gtgatcgttt ctgtatgggt ttgaattgaa
     5701 tcgcagaact gaagatcgat tggcattcct ggacagcacg tgctggtgct cacccgtttc
     5761 ctgcataggg acagctcatg gtgcacagct cagatcagat cgtgactcct cgagcggcgg
     5821 atgctggcga actgatctcc gccagcggac cggagatgag accccagcga accgataaca
     5881 gagcgagaga gctccagttc cgactgattg cacagtcggt gatctgggcg atgggcactg
     5941 ccagataggc tgggaattat caatcacttg aggtgaaagt gcggcgcaca caaatccaag
     6001 cttgatatca tcgatctcga ggctgcatcc aacgcgttgg gagctctccg gatcaattcg
     6061 gcttcaggta ccgtcgacga tgtaggtcac ggtctcgaag ccgcggtgcg ggtgccaggg
     6121 cgtgcccttg ggctccccgg gcgcgtactc cacctcaccc atctggtcca tcatgatgaa
     6181 cgggtcgagg tggcggtagt tgatcccggc gaacgcgcgg cgcaccggga agccctcgcc
     6241 ctcgaaaccg ctgggcgcgg tggtcacggt gagcacggga cgtgcgacgg cgtcggctgg
     6301 tgcggatacg cggggcagcg tcagcgggtt ctcgacggtc acggcgggca tgtcgacaag
     6361 ccgaattgat
//
