LOCUS       sequence-274768-        7469 bp    DNA     circular SYN 15-APR-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 7469)
  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 7469)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pHD-DsRed-kkv-3::NG
FEATURES             Location/Qualifiers
     source          1..7469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1086..1796
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1341..1350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2413..2446
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2672..2681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2678..3355
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(2808..2826)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2858..2878)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3076..3095
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3270..3293
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3476..3597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3513..3532)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3567..3586
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    3604..3637
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5276..5293)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5276..5292)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5391..5408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5562..6150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5642..5661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6272..7063,7064..7132))
                     /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     6895..6914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7133..7224)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7266..7285
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7291..7310
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttcgcccggg cgtattactc atcccccata ggtgcacatc tccaaggagt accttcagct
       61 ggagcccatc ggattggtgt tcgtattctt ctttgctctt attttgataa tccagttcac
      121 ggccatgctg ttccatcgat ttggaaccat ttcgcacatc ctggcctcga ctgaactcaa
      181 cttctgcaag aagaagtctg aggatctgac acaggaccaa ctaatagata aggtaggtgg
      241 cccgtaattg acattaatac aaataataaa attatgatat tatttagcat gctgtggaga
      301 tcgtcaagaa cttgcagagg ctgcagggaa tcgatggcga ctacgacaat gactccggtt
      361 ccgggccaga tcgcatagcc aggcggaaga ccattcaaaa tctggagaag gcgcgacagc
      421 cacgtcgcca gatcggcacc ctggatgtgg cgttcaagaa gcgcttcctg aaactcaccg
      481 ccgatgcgga gaacaatccg gccacgccca ttctcacccg ccgcctgaca atgcgggcgg
      541 agaccatccg ggcactggag gtgcgtaaga attcggtgat ggccgaaagg cggaagtccg
      601 cgatgcaaac tctgggagca aagaacgagt acggaatcac aactggagca ccggtgagat
      661 tagaaaagtt tttggagaaa ttttagatac aggatcataa cttctttaat gcatatatgt
      721 agattaacaa caatggagct ctgccgaatc agcgaagcgg acgtgtctcg aatgcaggca
      781 tcagcatcaa ggatgtattc aatgtgaacg gtggtggtgc cgaggtaaag gccaatttat
      841 aatcataaaa tatgtgtatt ctcatggaaa tattttgtta cgtccgctta cacttgcagc
      901 agatttacgg ctcgaatggg ggcggcacta tcaaccaggg ttacgagcat gtgatcgacg
      961 aggacggcga cggcaactcc ctgcggctga ccaccaggaa tccgcaccca cacccgcacc
     1021 accaggtctc ctggggccag aacaccaacg gcggcggcgg taatggcaca ggtcgcctgg
     1081 ccggcatggt gagcaagggc gaggaggata acatggcctc tctcccagcg acacatgagt
     1141 tacacatctt tggctccatc aacggtgtgg actttgacat ggtgggtcag ggcaccggca
     1201 atccaaatga tggttatgag gagttaaacc tgaagtccac caagggtgac ctccagttct
     1261 ccccctggat tctggtccct catatcgggt atggcttcca tcagtacctg ccctaccctg
     1321 acgggatgtc gcctttccag gccgccatgg tagatggctc cggataccaa gtccatcgca
     1381 caatgcagtt tgaagatggt gcctccctta ctgttaacta ccgctacacc tacgagggaa
     1441 gccacatcaa aggagaggcc caggtgaagg ggactggttt ccctgctgac ggtcctgtga
     1501 tgaccaactc gctgaccgct gcggactggt gcaggtcgaa gaagacttac cccaacgaca
     1561 aaaccatcat cagtaccttt aagtggagtt acaccactgg aaatggcaag cgctaccgga
     1621 gcactgcgcg gaccacctac acctttgcca agccaatggc ggctaactat ctgaagaacc
     1681 agccgatgta cgtgttccgt aagacggagc tcaagcactc caagaccgag ctcaacttca
     1741 aggagtggca aaaggccttt accgatgtga tgggcatgga cgagctgtac aagtaagttt
     1801 aaactgaagg acccgcccat cctagacgta gagctaagtt agacctaggt aacttgttaa
     1861 cgaattccgc ctctttccat caactgccta aacttaagac taagccaata cctagaattg
     1921 tagtcagaag gaaggattgt gttgtatagt atgtataggg acgcgagtga gagccaggac
     1981 actctggctc agaaggatca ggtcgttcat aaggcttcat ggcaacagaa cgaacattat
     2041 gcttagctaa tcacactgcc agtgtcgatg tgtgccaata attgggtggc tgatcgccca
     2101 ctgtggcagc aacattcatg cgatgtacta tactatcatt tgttaggtta tctttaagtc
     2161 aatacccgcg tggatcatcc gaaacacgcg gccagtgcat gctgacacaa aatactcaac
     2221 gtggagctga accgcagtgg cctatttgta cactcgacga catatcgagg atagttcttt
     2281 ttaaatgaat aatcattgat gatttgatgg attgtaaaac tttttaaaac aataaataga
     2341 agcatcaaac agatattcga tatttctcac tcatcaagct acgcggccgc ggacatatgc
     2401 acacctgcga tcataacttc gtataatgta tgctatacga agttatcgta cgggatctaa
     2461 ttcaattaga gactaattca attagagcta attcaattag gatccaagct tatcgatttc
     2521 gaaccctcga ccgccggagt ataaatagag gcgcttcgtc tacggagcga caattcaatt
     2581 caaacaagca aagtgaacac gtcgctaagc gaaagctaag caaataaaca agcgcagctg
     2641 aacaagctaa acaatcggct cgaagccggt cgccaccatg gcctcctccg aggacgtcat
     2701 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
     2761 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2821 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg
     2881 ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc
     2941 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     3001 ccaggactcc tccctccagg acggctcctt catctacaag gtgaagttca tcggcgtgaa
     3061 cttcccctcc gacggccccg taatgcagaa gaagactatg ggctgggagg cgtccaccga
     3121 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa
     3181 ggacggcggc cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca
     3241 gctgcccggc tactactacg tggactccaa gctggacatc acctcccaca acgaggacta
     3301 caccatcgtg gagcagtacg agcgcgccga gggccgccac cacctgttcc tgtaggggcc
     3361 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3421 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3481 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3541 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacc
     3601 ggtataactt cgtataatgt atgctatacg aagttataga agagcactag taaagatctc
     3661 gatatttctc actctcatca agcatcggtc atcccaagga taagctacat ccgaacccac
     3721 cttcggtagt gggataagct acatccgaac ccaccttcgg tagtggagac cctaccccaa
     3781 gtttagacca atacccgcat attttcggct tagagccctg gtactatacg aataacttta
     3841 ttttcgggaa agggtagaaa cattactaga catcgcttgc gggacaacat tgataaccag
     3901 ctatgataga taatagtatt agacacccat atggaaacgt cgtgtatgct cgcgttatca
     3961 gcgagttacc ttagtggaat aaaaggttgc ccttcttggc ggctatgggc aagcggtcgc
     4021 cctggaggaa cgggtttatg accgcgtcgt ccagccgagg atcgggcatc acaggatatc
     4081 cggccttctt agccaggctg ttaacgttgt tcaggctgtt ctcctggccc atctgcatga
     4141 tgtactgctc cacggcggag ttcaggtcac ccttgtcgcc gcccacatca gcactatcct
     4201 gttccgagca gtcgaacata tgctcgttgt cgcaggtaca gtcctgagcg gcctggaact
     4261 cccggctgaa gatggccgtg ttttcgaagt cgacaatgca gctgccgcct tgcgtctcca
     4321 tgtcgtagcc ctcagggcag ggattaggag ggttgcagta ggcaggcagg ccggcgtcgt
     4381 tcttcactat agtgggatac ctgttgggac cctctcccat tccgccggag atgaactggt
     4441 gaccccacag cgaactgtgc tgcagaaact cgtcatcccg cagcgaagga tgggaggcag
     4501 ctcccgatgc cgagcccgat tgcaggatgc agtcgcttcc gccgtcacac agcctcgcgt
     4561 actccgaccg ggagaccagg tccacgttgt ccttggagcc agcggctgcc tcgttgccca
     4621 cgtcgtaata gcccacctgc atgtctttgt ccatgcgatt cagcagatcc gtcatcaaga
     4681 catcggcaag aatatccttg gcgctatagg actgcacctg gtagccactc aaggcgacca
     4741 cgagcatcgc caccatcatg tacacacctg gaaaaacatg ggctattctc ttacctcgag
     4801 gctcttccgt caatcgagtt caagggcgac acaaaattta ttctaaatgc ataataaata
     4861 ctgataacat cttatagttt gtattatatt ttgtattatc gttgacatgt ataattttga
     4921 tatcaaaaac tgattttccc tttattattt tcgagattta ttttcttaat tctctttaac
     4981 aaactagaaa tattgtatat acaaaaaatc ataaataata gatgaatagt ttaattatag
     5041 gtgttcatca atcgaaaaag caacgtatct tatttaaagt gcgttgcttt tttctcattt
     5101 ataaggttaa ataattctca tatatcaagc aaagtgacag gcgcccttaa atattctgac
     5161 aaatgctctt tccctaaact ccccccataa aaaaacccgc cgaagcgggt ttttacgtta
     5221 tttgcggatt aacgattact cgttatcaga accgcccagg gggcccgagc ttaagactgg
     5281 ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa aaaggccatc cgtcaggggc
     5341 cttctgctta gtttgatgcc tggcagttcc ctactctcgc cttccgcttc ctcgctcact
     5401 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5941 aggtatgtag gcggtgctac agagttcttg aagtggtggg ctaactacgg ctacactaga
     6001 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6181 gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg gattttggtc atgagcttgc
     6241 gccgtcccgt caagtcagcg taatgctctg cttaccaatg cttaatcagt gaggcaccta
     6301 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6361 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     6421 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6481 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6541 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     6601 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6661 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6721 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6781 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6841 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6901 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6961 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7021 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7081 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     7141 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7201 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     7261 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg tttgcctggc
     7321 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     7381 gccgatggta gtgtggggac tccccatgcg agagtaggga actgccaggc atcaaataaa
     7441 acgaaaggct cagtcgaaag actgggcct
//
