LOCUS       sequence-273953-        6157 bp    DNA     circular SYN 14-APR-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 6157)
  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 6157)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pXF89
FEATURES             Location/Qualifiers
     source          1..6157
                     /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(3045..3064)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3046..3064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3071..3088)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3071..3087)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3080..3102)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3225..3527
                     /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     3406..3425
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3876..4670
                     /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             3876..4670
                     /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(3930..3949)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4540..4559
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      4870..4879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4876..5250
                     /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      5388..5976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5877..5896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6130..6147
                     /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 ctcttcgacc gccgcttcct
      361 ccggagatct cgagggtgga gagccaacgg gcgatctcgg aaccgatgag gtctgaaaaa
      421 ttcttttttt attgattttt agcattgaaa acatgaattt cagttaaaaa tctttttgaa
      481 aaatggaaat atactggaaa acggtcccga aaattgggga aagattttta gaacacataa
      541 tgtgaataaa aagctggaat tttaaatttt tcaaccaaaa tttagaaaaa gtatgaaaca
      601 atactttttc ataattttat tttttaattt acactttaca caatttaaac atattttttt
      661 aatttttaga aacatttttt gcattgaaaa atgtaccttt tcaaacaaat atacattttt
      721 caatatttta atttttttta ctttcatttt ttgtgtaatt aaaagcatct taaatttttt
      781 tttgcatttt tttaaacctt ttttttactg ttttggaaac attcttaggc gctttttttg
      841 ttaattttac atttaaaaat gctcttaaat atttttattc catttttcag ttcttaaact
      901 ttttttcaaa atctatgaac ttacctgggt tgtcctcttg gaggaggttg agtcctggtc
      961 cgatgtcgac ggccttgcag tttgggaggg acttggcgag acgggcggcc tcggctggtg
     1021 ggatgaggac tcctgaaaat tacggaaccg tattttttac gcaaaaaacc gcaaattttc
     1081 gaattttagt atcgtttttc gtaatataca atttgatttg tttctcttta taaataaaat
     1141 tacaaagaat tgcaggtttg aaaaaaacac gactttttta attaaaattt tctgttacat
     1201 ttttcaaatt tttttttgct tttttttttt gcaatttaac atttaaaaac tgtctctaaa
     1261 catattctta caacattttt taaatttgtt taaacattaa atattttttg cattaaaaaa
     1321 ctttgcattt tttcaaactt tttgaatttt taaatgttta ttttgttcaa ttttttagaa
     1381 attttatttt tattctaagc atttttgaaa acattttttt gcgttttttt tttaatttta
     1441 caaactacaa attagtacag ttttcggatt ttgtcccaaa atatttaaaa attgatgcgt
     1501 tttttaaaaa gaaaataaat gaaaattaga ctacaaactg cacttttcaa aaaaagaact
     1561 tacctggggt tccccagaag aggagctttg ggactgggga ttggtggagc cagtccatgt
     1621 actcctcgac gagggcgacg atgttggctg gctctccggc gattgggagc tcgtttggga
     1681 aacgccagag tggctcacgg tcgactgggt tgaggaatgg ctcacggtag tggtccatct
     1741 cgacctcggt gagtggacgg acgactccca ttgggagggt tccctcgatg aagacgtttt
     1801 ggtcgatgat gagcttacgt ccgacgtcgg tggtacggaa ggcttggaat gtctcacggg
     1861 cgaactctga aaatgttaaa tttgtagtta ttgtagttat tagattcaaa aaaagtaatt
     1921 gtataaatgt tttggtccaa aatttacaag ttttgtgaat tttctagttg aaaattgaaa
     1981 atttgaaaat tacatttaaa aaattgtgta tttctcaaaa cttttttgta tgaaattttt
     2041 tctcaaataa tgacttttgg ggctaaaaag catgaaaatt cacaatttta agggaaaaaa
     2101 tcgtatgaaa attgtttttt tttctggatt ttttgaagaa acttcacaaa aaatttaggc
     2161 ggtaggcagg caggtagaca taacttacct ggccactcgt cccaggttgg gattggacgg
     2221 atgaactcca tgaaggcgat tcccttgaca cgctctgggt tacgcttggc ccagtggaat
     2281 ccgagagccg atccccagtc gtggatgacg aggacgacct cctcgagtcc gagggcctcg
     2341 atgaaggcgt ccatgaaacg gacgtggtcg tcgaagaagt atccgaggtc tggcttgtcg
     2401 gactttccca ttccgatgag gtctgaaaat ctatttgaaa aatttacgag gaaattgttt
     2461 tttgtactaa attttcagga aaatctacaa tttgagccga aaaggtgaaa ttttatttaa
     2521 aaactgcaaa atttggggtt aaaatctggc aattctgaag aaaattcgct taaaaacgcg
     2581 atttcaacgt agtttttgat tttttttttt tttgttttac agcgtgaaat aatgaatatt
     2641 caacaatttt tggtgaaaat cccgcgaaaa attcaatttt tcaagtaaaa tctcggtttt
     2701 ttccttcttt cccataagaa aacttacctg gggcgatgca acggtgggtt ggggcgacgt
     2761 gtgggatgat gttacgccag acgtaggagg aggttgggtt tccgtggagg aagaggactg
     2821 gggttccgtc acgtggtccg acgtcgacgt agtgcatacg ctctccgagg acctcgacgt
     2881 agtgtgggtc gaatgggaat ccggttccga tcttaaaatt aatttttaaa cagagcaaca
     2941 aaaactaagt gttacctcgg ccatcgaaga gctgcaaggg cgaattctgc agatatccat
     3001 cacactggcg gccgctcgag catgcatcta gagggcccaa ttcgccctat agtgagtcgt
     3061 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3121 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3181 gcaccgatcg cccttcccaa cagttgcgca gcctatacgt acggcagttt aaggtttaca
     3241 cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca
     3301 cgccggggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct
     3361 cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg
     3421 atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg
     3481 aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggcatga
     3541 gattatcaaa aaggatcttc acctagatcc ttttcacgta gaaagccagt ccgcagaaac
     3601 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     3661 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     3721 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     3781 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     3841 gctctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3901 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3961 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4021 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4081 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4141 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4201 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4261 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4321 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4381 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4441 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4501 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4561 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4621 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attattaacg
     4681 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4741 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4801 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatagcac
     4861 gtgaggaggg ccaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     4921 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     4981 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     5041 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     5101 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     5161 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     5221 gtgcacttcg tggccgagga gcaggactga cacgtgctaa aacttcattt ttaatttaaa
     5281 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5341 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5401 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5461 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5521 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5581 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5641 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5701 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5761 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5821 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5881 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5941 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6001 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6061 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6121 accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
