LOCUS       sequence-335013-        2560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multiple cloning site part for enhancer cloning, requires minimal 
            promoter element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2560)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 2560)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pMCS-TCCC
FEATURES             Location/Qualifiers
     source          1..2560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..35)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    68..124
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(183..201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(242..260)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(276..292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(276..292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(407..995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(487..506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1110..1212
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1213..1872
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1279..1298)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2459..2476
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2460..2476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2483..2501
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        join(2549..2560,1..7)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 ctatagactt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgtcgaag
       61 tcttctcaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
      121 attcgaagac ttgcttgtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      181 ggcctatagt gagtcgtatt atccccgaga ccactcatcg cccaatcact agtgaattcg
      241 ttctatagtg tcacctaaat accatagagc cgttggtcat agctgtttcc tgtccaggca
      301 tcatgacacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      361 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      421 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      481 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      541 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      601 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      661 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      721 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      781 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      841 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      901 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      961 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1021 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1081 aaaggatctt cacctagatc cttttgtgat gatcggcacg taagaggttc caactttcac
     1141 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     1201 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     1261 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     1321 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     1381 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca
     1441 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     1501 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     1561 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     1621 gggtttattg agaatatgtt tttcgtgtca gccaatccct gggtgagttt caccagtttt
     1681 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     1741 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     1801 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     1861 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg ttgctacgcc
     1921 tgaataagtg ataataagcg gatgaatggc agaaattcga tgataagctg tcaaacatga
     1981 tgttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccagcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt taacattgta gaaacgcatg
     2461 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggtgaattcc gcttaggacc
     2521 ttagatattg atctcaaggt ctcgggagat ttaggtgaca
//
