LOCUS       sequence-273961-        7136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MosSCI SapTrap assembly: Cassette 3 for 4-cassette system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7136)
  AUTHORS   Fan X, Henau S, Feinstein J, Miller SI, Han B, Frokjaer-Jensen C, 
            Griffin EE
  TITLE     SapTrap Assembly of Caenorhabditis elegans MosSCI Transgene Vectors.
  JOURNAL   G3 (Bethesda). 2019 Dec 17. pii: g3.119.400822. doi: 
            10.1534/g3.119.400822.
  PUBMED    31848219
REFERENCE   2  (bases 1 to 7136)
  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: "G3 
            (Bethesda). 2019 Dec 17. pii: g3.119.400822. doi: 
            10.1534/g3.119.400822."
COMMENT     Sequence Label: pJF6
FEATURES             Location/Qualifiers
     source          1..7136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4024..4043)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4025..4043)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4050..4067)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4050..4066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4059..4081)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4204..4506
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4385..4404
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4855..5649
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4855..5649
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4909..4928)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5519..5538
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      5849..5858
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5855..6229
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      6367..6955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6856..6875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7109..7126
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttgcag ctcttcgggt ggaagcggct
      361 ccaaggtaac acttagtttt tgttgctctg tttaaaaatt aattttaagg gagaggagct
      421 gttcaccgga gtcgtcccaa tcctcgtcga gctcgacggg gacgtcaacg gacacaagtt
      481 ctccgtgtcc ggagagggag agggagatgc cacctacgga aagctcaccc tcaagttcat
      541 ctgcaccacc ggaaagctcc cagtcccatg gccaaccctc gtcaccacct tctgctacgg
      601 agtccaatgc ttctcccgtt acccaggtaa gttttctgtt ttatttactt gttttgagaa
      661 ttgggatttt tagctgaaaa attggatttt aagatgaaaa attgaaattt tcaactaaaa
      721 atttcaattt ttagctgaaa agttaaattt ttagctaaag agtgggattt ttagttgaaa
      781 aatcgagatt tctagctaaa aagtatgatt tttagctgaa agattggaat tttaagcaga
      841 aaaaattgta tttttagctg ataaatcaaa ttttaagcgg aaaaatttga atttttagct
      901 aaaaaattcc attttcaact aaaaatcgtg atttttagct ttaaaaattg gatttttagc
      961 tgaaaaatcg gatttttagc tgaaaattga aattttttag ttaaaaagtg agatttttta
     1021 acagaaaagt gagattttta gttaaaaaat aatattttta gctaaaaaat acaattttca
     1081 actaaagatc gtgattttta gctttaaaaa ttggattttt agctaaaaaa tcggattttt
     1141 agctgaaaat tgaaattttt tagttaaaaa gtgggatttt aagatgaaaa gtgagatttt
     1201 ttagctgaaa agtgagattt tcagctaaaa aatacaattt tcaactgaaa aattgggatt
     1261 tttagcttta aaaattggat tttcagcttt aaaaattgga tttttagctg aaaagtgaga
     1321 tttttagttg aaaaattgaa atgtttagct gaaaaattgg aatttttaag taaaaaagtg
     1381 ggattttaag atgaaaagtg agatgttttt agctgaaaag tgagattttt agctaaaaaa
     1441 tacaattttc aactgaaaaa ttgggatttt tagctcaaaa gtgggacgaa agacgaaaaa
     1501 aaaaatcgat tttttaccaa aaattagatt ttcagaccac atgaagcgtc acgacttctt
     1561 caagtccgcc atgccagagg gatacgtcca agagcgtacc atcttcttca aggacgacgg
     1621 aaactacaag gtaagttatg tacgcgattt ctcggcgaaa aaatggactt ttttgctcaa
     1681 aaactgaatt tttcagcttt ttttgcactg aaaatgctga aaattggttt ttttgctgaa
     1741 aattcaccga aaaaagctga aaatattgca gaaatcgacg aaaaatgcct tttttgctga
     1801 aatttcagcc agaaaattgt tcagaatcgc aatttcctgc caaaatcgac gtttttcggc
     1861 gaaaattggt agattttgaa ccgaaaattg tgaaaaatgg acttttttgc tcaaaaacag
     1921 actttttgag cttttttcac actgaaaatg gtggaaaatt ggattttcca gcagaaaatt
     1981 cggaaatttt caatttttgg cggaaatttg acgaaaaata tatgaaaaac accgattttt
     2041 gtctgaaaaa gttcaaaagc cgaaaaattg gccaaaaatc tccaaaatcg gacttttctg
     2101 gcgtttttcg tggaaaaatt gaaatttttt tgaaaaattg agcaaaaatc gccgaaaaac
     2161 cgctaaaaaa actgccgaaa ttggcaattt tgagcttaaa aaaacttgaa aaatgctcaa
     2221 aattacccaa ttttcaccgg aaattgagct aaaatcgtcg aaaaagactg aattttcaat
     2281 ttttagctga aaatctgcct attttcggct aaaattgctt taaaaacatt aaaattcaaa
     2341 ttttttggct aaaaatactc agattgccct tttttttctc caaaaatgac aaaaaaaaaa
     2401 tcaatcagaa aaatgcgatt ttttttacaa aaattggctt caaaacgtct aaattctccc
     2461 taatttttgc tgctgaaaat tgtgcaaaaa tcaccgaaaa atgacgattt taggctgaaa
     2521 atcttcaaaa aaagctcaaa attccccact tttcaccgga aaaagacgaa aaaagctcca
     2581 aatttatcga ttaacatttt cagacccgtg ccgaggtcaa gttcgaggga gacaccctcg
     2641 tcaaccgtat cgagctcaag ggaatcgact tcaaggagga cggtaacatc ctgggacaca
     2701 agctggagta caactacaac tcccacaacg tctacatcat ggccgacaag caaaagaacg
     2761 gaatcaaggt caacttcaag gtaagttctt ttttttgctc gtttttgagt gaaaaattga
     2821 tggaattcac aaaaaatggc caaagaatcc tcaaaattgg ccgaaaattc tgaaaaatga
     2881 cctaaaaatc accgaaaaac ggccaaaaat tatctgaaaa tcgccgaaaa tctcaaattt
     2941 tgaggaaatt cccgctaaaa attaataaaa atcaccgaaa aaaggccaaa aattacctaa
     3001 taaaggccga aaatcgccta aaaaggccaa aaattctgaa aaatggccaa atttcaattt
     3061 ttcacacaca aaaaatccct gaattttcta tttttactcg aaaaactgtg gatttacggg
     3121 ggtaaaaatt cagtttttag tgctatttca caagaaaaac acagcatttt cgacaaaaac
     3181 tccgattttt aagccaaaaa tttgaggttt ttagtttttt acgtgaaaac ctgaactttt
     3241 tgccaaaaaa acactttttt ttagactaaa aatcaatttt cgatcaaaaa tcactaattt
     3301 tgcctgaaaa aatatagatt tttccacttt ttcactgaaa aaaaactgga ttttcactag
     3361 aaaatcaagt tgtttttaga ccaaaaattt tatttttccg ctgaaaatta tcgtttttca
     3421 gtcaattttc agctaaaaat ccctgaaatt ctggattttc agccaaaaaa ctcgaaattt
     3481 tccaaaaaaa ccccgtttaa aaatgagttt agactgaaaa ttccgatttt tgagccaaaa
     3541 atttgatttt taatcgaaaa aaattggatt ttcaaccaaa aaatcagttt ttcagtgatt
     3601 tttagagtaa aaatcactga ttttaacaca aaattccagt ttttcgtgat ttttcactag
     3661 aaaatcccga tttttcccca aaaaatccat ttttccgact aaaaatcccc aaagtaattt
     3721 tcagatccgt cacaacatcg aggacggatc ggtccaactc gccgaccact accaacaaaa
     3781 caccccaatc ggtgacggac cagtcctcct cccagacaac cactacctct ccacccaatc
     3841 cgccctctcc aaggacccaa acgagaagcg tgaccacatg gtcctcctcg agttcgtcac
     3901 cgccgccgga atcacccacg gaatggacga gctctacaag taacgaagag ctgcaagggc
     3961 gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggcccaat
     4021 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     4081 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4141 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctatacgta
     4201 cggcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     4261 cagagtgata ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt
     4321 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     4381 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     4441 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     4501 atataaatgt caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag
     4561 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
     4621 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     4681 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     4741 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     4801 tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4861 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4921 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4981 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5041 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5101 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5161 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5221 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5281 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5341 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5401 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5461 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5521 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5581 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5641 ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     5701 tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5761 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5821 aatgcttcaa taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc
     5881 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     5941 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     6001 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     6061 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     6121 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     6181 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctaaa
     6241 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6301 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6361 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6421 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6481 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6541 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6601 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6661 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6721 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6781 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6841 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6901 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6961 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7021 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7081 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
