LOCUS       sequence-315468-        3084 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Confers resistance to hygromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3084)
  AUTHORS   Andreou , Andreas I., Nirkko, Jessica , Ochoa-Villarreal, Marisol , 
            Nakayama, Naomi
  TITLE     Mobius Assembly for Plant Systems highlights promoter-terminator 
            interaction in gene regulation
  JOURNAL   bioRxiv, March 31, 2021
REFERENCE   2  (bases 1 to 3084)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv, 
            March 31, 2021"
COMMENT     Sequence Label: lvl0 HptII
FEATURES             Location/Qualifiers
     source          1..3084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      21..78
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(102..119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(273..861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(353..372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1044..1138)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1159..1818)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1733..1752
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1980..1998
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2002..2045)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             join(2062..3084,1..3)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             join(2062..3084,1..3)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 taggctttga gaccctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt
       61 tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg
      121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      241 aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag
      301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
      961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1021 tggtctgaca gctcgaggct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     1081 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     1141 agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     1201 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     1261 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
     1321 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     1381 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     1441 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     1501 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     1561 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat
     1621 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     1681 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     1741 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     1801 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     1861 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat
     1921 caactcgagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     1981 ataggcgtat cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg
     2041 atttctggaa ttcggtctct aatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     2101 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     2161 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     2221 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     2281 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     2341 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     2401 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     2461 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     2521 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     2581 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     2641 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     2701 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     2761 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     2821 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     2881 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     2941 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     3001 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     3061 agcactcgtc cgagggcaaa ggaa
//
