LOCUS       sequence-327008-        3404 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 3404)
  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 3404)
  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..3404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     77..93
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     77..93
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(163..179)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(164..180)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(205..223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(233..250)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(233..249)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(242..264)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      391..846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(478..497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     688..709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1002..1661
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1068..1087)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1832..2420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2321..2340
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2613..2984)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3017..3126
                     /note="incP origin of transfer"
     primer_bind     3364..3381
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccggcag atctgatatc atcgatgaat tcgagctcca
       61 ccgcggtggc ggccgctcta gaactagtgg atccaccggt tccatggcta gccacgtgac
      121 gcgtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga
      181 gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
      241 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
      301 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
      361 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
      421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
      481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
      541 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
      601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
      661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
      721 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
      781 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
      841 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      901 aatacaacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
      961 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     1021 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     1081 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     1141 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     1201 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     1261 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1321 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1381 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     1441 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1501 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     1561 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     1621 aattacaaca gtactgcgat gagtggcagg gcggggcgta actgtcagac caagtttact
     1681 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1741 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1801 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1861 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1921 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1981 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2041 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2101 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2161 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2221 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2281 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2341 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2401 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2461 gctggccttt tgctcacatg tggcgctcgg tcttgccttg ctcgtcggtg atgtacttca
     2521 ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg gacgtgcttg gcaatcacgc
     2581 gcaccccccg gccgttttag cggctaaaaa agtcatggct ctgccctcgg gcggaccacg
     2641 cccatcatga ccttgccaag ctcgtcctgc ttctcttcga tcttcgccag cagggcgagg
     2701 atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg tgagccagag tttcagcagg
     2761 ccgcccaggc ggcccaggtc gccattgatg cgggccagct cgcggacgtg ctcatagtcc
     2821 acgacgcccg tgattttgta gccctggccg acggccagca ggtaggccga caggctcatg
     2881 ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca gtacaccttg
     2941 ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt gccctcatct
     3001 gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
     3061 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     3121 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     3181 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     3241 cagggttatg cagcggaaaa gcgctacatg ttctttcctg cgttatcccc tgattctgtg
     3301 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3361 cgcagcgagt cagtgagcga ggaagcaacc tggcttatcg aaat
//
