LOCUS       sequence-249814-        3497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes pCMV-Hygromycin resistance cassette as a type 6 part to be 
            used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3497)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 3497)
  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"; doi: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK6_009
FEATURES             Location/Qualifiers
     source          1..3497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      98..686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     587..606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1132..1335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1286..1306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1357..1366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1363..2391
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1363..2391
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     primer_bind     complement(2403..2420)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2409..2616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      complement(2649..2683)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2754..3413)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3328..3347
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 acctcttacg tgcccgatca atcatgacca aaatccctta acgtgagttt tcgttccact
       61 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      121 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      181 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      241 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      301 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      361 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      421 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      481 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
      541 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      601 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      661 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
      721 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      781 accgtagtcg gtctcataca gcggtgggct ctatggggta cctacgtagg tacataactt
      841 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      901 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      961 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     1021 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1081 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1141 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1201 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1261 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     1321 tatataagca gagctcgttt agtgaaccgt cagatagcca ccatgggtaa aaagcctgaa
     1381 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt gtccgacctg
     1441 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     1501 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     1561 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga atttagcgag
     1621 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     1681 accgaactgc ccgctgttct gcaaccggtc gcggaggcca tggatgcgat cgctgcggcc
     1741 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     1801 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     1861 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     1921 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     1981 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     2041 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     2101 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     2161 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     2221 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     2281 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     2341 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata actcgagcga
     2401 cgcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2461 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2521 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2581 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tgagttgaga
     2641 ccagaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     2701 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc
     2761 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     2821 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     2881 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     2941 aaatcaaaac tggtgaaact cacccaggga ttggctgaaa cgaaaaacat attctcaata
     3001 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     3061 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     3121 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     3181 ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     3241 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     3301 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     3361 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     3421 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     3481 ttatggtgaa agttgga
//
