LOCUS       sequence-332642-        2927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  It is used to bypass the Level 1 cloning and directly feed 
            terminators into Level 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2927)
  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 2927)
  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: "bioRxiv, 
            March 31, 2021"
COMMENT     Sequence Label: lvl0 MethylAblet
FEATURES             Location/Qualifiers
     source          1..2927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      24..81
                     /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(105..122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(276..864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(356..375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1047..1141)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1162..1821)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1736..1755
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1983..2001
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2005..2048)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        2077..2111
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             2120..2131
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2811..2882
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2898..2925
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 gtctcgcgct tgagaccctg cagtccggca aaaaagggca aggtgtcacc accctgccct
       61 ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct cactgactcg
      121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac
      301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1021 acttggtctg acagctcgag gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     1081 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     1141 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     1201 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     1261 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     1321 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     1381 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     1441 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     1501 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     1561 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     1621 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     1681 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     1741 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     1801 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     1861 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc
     1921 gatcaactcg agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     1981 aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg
     2041 atgatttctg gaattcaggt ctctgcttcg agaccgttta cggctagctc agtcctaggt
     2101 atagtgctag ctactagaga aagaggagaa atactaaatg agtgtgatcg ctaaacaaat
     2161 gacctacaag gtttatatgt caggcacggt caatggacac tactttgagg tcgaaggcga
     2221 tggaaaaggt aagccctacg agggggagca gacggtaaag ctcactgtca ccaagggcgg
     2281 acctctgcca tttgcttggg atattttatc accacagtgt cagtacggaa gcataccatt
     2341 caccaagtac cctgaagaca tccctgacta tgtaaagcag tcattcccgg agggctatac
     2401 atgggagagg atcatgaact ttgaagatgg tgcagtgtgt actgtcagca atgattccag
     2461 catccaaggc aactgtttca tctaccatgt caagttctct ggtttgaact ttcctcccaa
     2521 tggacctgtc atgcagaaga agacacaggg ctgggaaccc aacactgagc gtctctttgc
     2581 acgagatgga atgctgctag gaaacaactt tatggctctg aagttagaag gaggcggtca
     2641 ctatttgtgt gaatttaaaa ctacttacaa ggcaaagaag cctgtgaaga tgccagggta
     2701 tcactatgtt gaccgcaaac tggatgtaac caatcacaac aaggattaca cttcggttga
     2761 gcagtgtgaa atttccattg cacgcaaacc tgtggtcgcc taataagctt caaataaaac
     2821 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2881 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgcg
//
