LOCUS       sequence-273915-        7136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MosSCI SapTrap assembly: Cassette 2 for 4- or 5-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: pJF5
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 ctcttcgatg tccaaggtaa
      361 cacttagttt ttgttgctct gtttaaaaat taattttaag ggagaggagc tgttcaccgg
      421 agtcgtccca atcctcgtcg agctcgacgg ggacgtcaac ggacacaagt tctccgtgtc
      481 cggagaggga gagggagatg ccacctacgg aaagctcacc ctcaagttca tctgcaccac
      541 cggaaagctc ccagtcccat ggccaaccct cgtcaccacc ttctgctacg gagtccaatg
      601 cttctcccgt tacccaggta agttttctgt tttatttact tgttttgaga attgggattt
      661 ttagctgaaa aattggattt taagatgaaa aattgaaatt ttcaactaaa aatttcaatt
      721 tttagctgaa aagttaaatt tttagctaaa gagtgggatt tttagttgaa aaatcgagat
      781 ttctagctaa aaagtatgat ttttagctga aagattggaa ttttaagcag aaaaaattgt
      841 atttttagct gataaatcaa attttaagcg gaaaaatttg aatttttagc taaaaaattc
      901 cattttcaac taaaaatcgt gatttttagc tttaaaaatt ggatttttag ctgaaaaatc
      961 ggatttttag ctgaaaattg aaatttttta gttaaaaagt gagatttttt aacagaaaag
     1021 tgagattttt agttaaaaaa taatattttt agctaaaaaa tacaattttc aactaaagat
     1081 cgtgattttt agctttaaaa attggatttt tagctaaaaa atcggatttt tagctgaaaa
     1141 ttgaaatttt ttagttaaaa agtgggattt taagatgaaa agtgagattt tttagctgaa
     1201 aagtgagatt ttcagctaaa aaatacaatt ttcaactgaa aaattgggat ttttagcttt
     1261 aaaaattgga ttttcagctt taaaaattgg atttttagct gaaaagtgag atttttagtt
     1321 gaaaaattga aatgtttagc tgaaaaattg gaatttttaa gtaaaaaagt gggattttaa
     1381 gatgaaaagt gagatgtttt tagctgaaaa gtgagatttt tagctaaaaa atacaatttt
     1441 caactgaaaa attgggattt ttagctcaaa agtgggacga aagacgaaaa aaaaaatcga
     1501 ttttttacca aaaattagat tttcagacca catgaagcgt cacgacttct tcaagtccgc
     1561 catgccagag ggatacgtcc aagagcgtac catcttcttc aaggacgacg gaaactacaa
     1621 ggtaagttat gtacgcgatt tctcggcgaa aaaatggact tttttgctca aaaactgaat
     1681 ttttcagctt tttttgcact gaaaatgctg aaaattggtt tttttgctga aaattcaccg
     1741 aaaaaagctg aaaatattgc agaaatcgac gaaaaatgcc ttttttgctg aaatttcagc
     1801 cagaaaattg ttcagaatcg caatttcctg ccaaaatcga cgtttttcgg cgaaaattgg
     1861 tagattttga accgaaaatt gtgaaaaatg gacttttttg ctcaaaaaca gactttttga
     1921 gcttttttca cactgaaaat ggtggaaaat tggattttcc agcagaaaat tcggaaattt
     1981 tcaatttttg gcggaaattt gacgaaaaat atatgaaaaa caccgatttt tgtctgaaaa
     2041 agttcaaaag ccgaaaaatt ggccaaaaat ctccaaaatc ggacttttct ggcgtttttc
     2101 gtggaaaaat tgaaattttt ttgaaaaatt gagcaaaaat cgccgaaaaa ccgctaaaaa
     2161 aactgccgaa attggcaatt ttgagcttaa aaaaacttga aaaatgctca aaattaccca
     2221 attttcaccg gaaattgagc taaaatcgtc gaaaaagact gaattttcaa tttttagctg
     2281 aaaatctgcc tattttcggc taaaattgct ttaaaaacat taaaattcaa attttttggc
     2341 taaaaatact cagattgccc ttttttttct ccaaaaatga caaaaaaaaa atcaatcaga
     2401 aaaatgcgat tttttttaca aaaattggct tcaaaacgtc taaattctcc ctaatttttg
     2461 ctgctgaaaa ttgtgcaaaa atcaccgaaa aatgacgatt ttaggctgaa aatcttcaaa
     2521 aaaagctcaa aattccccac ttttcaccgg aaaaagacga aaaaagctcc aaatttatcg
     2581 attaacattt tcagacccgt gccgaggtca agttcgaggg agacaccctc gtcaaccgta
     2641 tcgagctcaa gggaatcgac ttcaaggagg acggtaacat cctgggacac aagctggagt
     2701 acaactacaa ctcccacaac gtctacatca tggccgacaa gcaaaagaac ggaatcaagg
     2761 tcaacttcaa ggtaagttct tttttttgct cgtttttgag tgaaaaattg atggaattca
     2821 caaaaaatgg ccaaagaatc ctcaaaattg gccgaaaatt ctgaaaaatg acctaaaaat
     2881 caccgaaaaa cggccaaaaa ttatctgaaa atcgccgaaa atctcaaatt ttgaggaaat
     2941 tcccgctaaa aattaataaa aatcaccgaa aaaaggccaa aaattaccta ataaaggccg
     3001 aaaatcgcct aaaaaggcca aaaattctga aaaatggcca aatttcaatt tttcacacac
     3061 aaaaaatccc tgaattttct atttttactc gaaaaactgt ggatttacgg gggtaaaaat
     3121 tcagttttta gtgctatttc acaagaaaaa cacagcattt tcgacaaaaa ctccgatttt
     3181 taagccaaaa atttgaggtt tttagttttt tacgtgaaaa cctgaacttt ttgccaaaaa
     3241 aacacttttt tttagactaa aaatcaattt tcgatcaaaa atcactaatt ttgcctgaaa
     3301 aaatatagat ttttccactt tttcactgaa aaaaaactgg attttcacta gaaaatcaag
     3361 ttgtttttag accaaaaatt ttatttttcc gctgaaaatt atcgtttttc agtcaatttt
     3421 cagctaaaaa tccctgaaat tctggatttt cagccaaaaa actcgaaatt ttccaaaaaa
     3481 accccgttta aaaatgagtt tagactgaaa attccgattt ttgagccaaa aatttgattt
     3541 ttaatcgaaa aaaattggat tttcaaccaa aaaatcagtt tttcagtgat ttttagagta
     3601 aaaatcactg attttaacac aaaattccag tttttcgtga tttttcacta gaaaatcccg
     3661 atttttcccc aaaaaatcca tttttccgac taaaaatccc caaagtaatt ttcagatccg
     3721 tcacaacatc gaggacggat cggtccaact cgccgaccac taccaacaaa acaccccaat
     3781 cggtgacgga ccagtcctcc tcccagacaa ccactacctc tccacccaat ccgccctctc
     3841 caaggaccca aacgagaagc gtgaccacat ggtcctcctc gagttcgtca ccgccgccgg
     3901 aatcacccac ggaatggacg agctctacaa gggaagcggc ggtcgaagag 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
//
