LOCUS       sequence-273941-        6169 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 6169)
  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 6169)
  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: pXF90
FEATURES             Location/Qualifiers
     source          1..6169
                     /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(3057..3076)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3058..3076)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3083..3100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3083..3099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3092..3114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3237..3539
                     /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     3418..3437
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3888..4682
                     /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             3888..4682
                     /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(3942..3961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4552..4571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      4882..4891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4888..5262
                     /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      5400..5988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5889..5908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6142..6159
                     /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 ggaagcggcg
      361 ccgaggtaac acttagtttt tgttgctctg tttaaaaatt aattttaaga tcggaaccgg
      421 attcccattc gacccacact acgtcgaggt cctcggagag cgtatgcact acgtcgacgt
      481 cggaccacgt gacggaaccc cagtcctctt cctccacgga aacccaacct cctcctacgt
      541 ctggcgtaac atcatcccac acgtcgcccc aacccaccgt tgcatcgccc caggtaagtt
      601 ttcttatggg aaagaaggaa aaaaccgaga ttttacttga aaaattgaat ttttcgcggg
      661 attttcacca aaaattgttg aatattcatt atttcacgct gtaaaacaaa aaaaaaaaaa
      721 atcaaaaact acgttgaaat cgcgttttta agcgaatttt cttcagaatt gccagatttt
      781 aaccccaaat tttgcagttt ttaaataaaa tttcaccttt tcggctcaaa ttgtagattt
      841 tcctgaaaat ttagtacaaa aaacaatttc ctcgtaaatt tttcaaatag attttcagac
      901 ctcatcggaa tgggaaagtc cgacaagcca gacctcggat acttcttcga cgaccacgtc
      961 cgtttcatgg acgccttcat cgaggccctc ggactcgagg aggtcgtcct cgtcatccac
     1021 gactggggat cggctctcgg attccactgg gccaagcgta acccagagcg tgtcaaggga
     1081 atcgccttca tggagttcat ccgtccaatc ccaacctggg acgagtggcc aggtaagtta
     1141 tgtctacctg cctgcctacc gcctaaattt tttgtgaagt ttcttcaaaa aatccagaaa
     1201 aaaaaacaat tttcatacga ttttttccct taaaattgtg aattttcatg ctttttagcc
     1261 ccaaaagtca ttatttgaga aaaaatttca tacaaaaaag ttttgagaaa tacacaattt
     1321 tttaaatgta attttcaaat tttcaatttt caactagaaa attcacaaaa cttgtaaatt
     1381 ttggaccaaa acatttatac aattactttt tttgaatcta ataactacaa taactacaaa
     1441 tttaacattt tcagagttcg cccgtgagac attccaagcc ttccgtacca ccgacgtcgg
     1501 acgtaagctc atcatcgacc aaaacgtctt catcgaggga accctcccaa tgggagtcgt
     1561 ccgtccactc accgaggtcg agatggacca ctaccgtgag ccattcctca acccagtcga
     1621 ccgtgagcca ctctggcgtt tcccaaacga gctcccaatc gccggagagc cagccaacat
     1681 cgtcgccctc gtcgaggagt acatggactg gctccaccaa tccccagtcc caaagctcct
     1741 cttctgggga accccaggta agttcttttt ttgaaaagtg cagtttgtag tctaattttc
     1801 atttattttc tttttaaaaa acgcatcaat ttttaaatat tttgggacaa aatccgaaaa
     1861 ctgtactaat ttgtagtttg taaaattaaa aaaaaaacgc aaaaaaatgt tttcaaaaat
     1921 gcttagaata aaaataaaat ttctaaaaaa ttgaacaaaa taaacattta aaaattcaaa
     1981 aagtttgaaa aaatgcaaag ttttttaatg caaaaaatat ttaatgttta aacaaattta
     2041 aaaaatgttg taagaatatg tttagagaca gtttttaaat gttaaattgc aaaaaaaaaa
     2101 agcaaaaaaa aatttgaaaa atgtaacaga aaattttaat taaaaaagtc gtgttttttt
     2161 caaacctgca attctttgta attttattta taaagagaaa caaatcaaat tgtatattac
     2221 gaaaaacgat actaaaattc gaaaatttgc ggttttttgc gtaaaaaata cggttccgta
     2281 attttcagga gtcctcatcc caccagccga ggccgcccgt ctcgccaagt ccctcccaaa
     2341 ctgcaaggcc gtcgacatcg gaccaggact caacctcctc caagaggaca acccaggtaa
     2401 gttcatagat tttgaaaaaa agtttaagaa ctgaaaaatg gaataaaaat atttaagagc
     2461 atttttaaat gtaaaattaa caaaaaaagc gcctaagaat gtttccaaaa cagtaaaaaa
     2521 aaggtttaaa aaaatgcaaa aaaaaattta agatgctttt aattacacaa aaaatgaaag
     2581 taaaaaaaat taaaatattg aaaaatgtat atttgtttga aaaggtacat ttttcaatgc
     2641 aaaaaatgtt tctaaaaatt aaaaaaatat gtttaaattg tgtaaagtgt aaattaaaaa
     2701 ataaaattat gaaaaagtat tgtttcatac tttttctaaa ttttggttga aaaatttaaa
     2761 attccagctt tttattcaca ttatgtgttc taaaaatctt tccccaattt tcgggaccgt
     2821 tttccagtat atttccattt ttcaaaaaga tttttaactg aaattcatgt tttcaatgct
     2881 aaaaatcaat aaaaaaagaa tttttcagac ctcatcggtt ccgagatcgc ccgttggctc
     2941 tccaccctcg agatctccgg acatgacgaa ctctaacgaa gagctgcaag ggcgaattct
     3001 gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc aattcgccct
     3061 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3121 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3181 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctatac gtacggcagt
     3241 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     3301 atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     3361 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     3421 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     3481 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     3541 tgtcaggcat gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca
     3601 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     3661 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     3721 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     3781 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     3841 gcaggggatc aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3901 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3961 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4021 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4081 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4141 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4201 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4261 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4321 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4381 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4441 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4501 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4561 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4621 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4681 gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4741 tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4801 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4861 caataatagc acgtgaggag ggccaccatg gccaagttga ccagtgccgt tccggtgctc
     4921 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     4981 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     5041 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     5101 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     5161 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     5221 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct aaaacttcat
     5281 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5521 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
