LOCUS       sequence-317220-        3394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified L4440 containing now CamR and oriT for conjugation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3394)
  AUTHORS   Kazmierczak M, Farre I Diaz C, Ofenbauer A, Herzog S, Tursun B
  TITLE     The CONJUDOR pipeline for multiplexed knockdown of gene pairs 
            identifies RBBP-5 as a germ cell reprogramming barrier in C. 
            elegans.
  JOURNAL   Nucleic Acids Res. 2021 Feb 26;49(4):e22. doi: 10.1093/nar/gkaa1171.
  PUBMED    33290523
REFERENCE   2  (bases 1 to 3394)
  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: "Nucleic 
            Acids Res."; date: "2021-02-26"; volume: "49(4):e22. doi"; pages: " 
            10.1093/nar/gkaa1171"
COMMENT     Sequence Label: LoriT
FEATURES             Location/Qualifiers
     source          1..3394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      354..942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     843..862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1132..1503)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1536..1645
                     /note="incP origin of transfer"
     primer_bind     1880..1897
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1921..1939
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1997..2013
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1997..2013
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2121..2139)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2149..2166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2149..2165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2158..2180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2307..2762
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2394..2413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2604..2625
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(2918..3394,1..183)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2984..3003)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
       61 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
      121 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
      181 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      241 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      301 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      361 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      421 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      481 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
      541 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      601 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      661 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      721 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      781 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      841 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      901 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      961 tttttacggt tcctggcctt ttgctggcct tttgctcaca ggcgctcggt cttgccttgc
     1021 tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga gcgcatgggg
     1081 acgtgcttgg caatcacgcg caccccccgg ccgttttagc ggctaaaaaa gtcatggctc
     1141 tgccctcggg cggaccacgc ccatcatgac cttgccaagc tcgtcctgct tctcttcgat
     1201 cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt
     1261 gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc gggccagctc
     1321 gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga cggccagcag
     1381 gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc
     1441 tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg tttcatcagc
     1501 catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca gagcaggatt
     1561 cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg
     1621 tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac
     1681 acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat
     1741 cgctataatg accccgaagc agggttatgc agcggaaaag cgcttgttct ttcctgcgtt
     1801 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     1861 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcaacctggc ttatcgaaat
     1921 taatacgact cactataggg agaccggcag atctgatatc atcgatgaat tcgagctcca
     1981 ccgcggtggc ggccgctcta gaactagtgg atccaccggt tccatggcta gccacgtgac
     2041 gcgtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg cctcgagggg
     2101 gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     2161 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2221 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2281 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     2341 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2401 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2461 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2521 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2581 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2641 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2701 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     2761 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2821 caacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     2881 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     2941 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     3001 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     3061 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     3121 tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     3181 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     3241 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     3301 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     3361 tccctgggtg agtttcacca gttttgattt aaac
//
