LOCUS       sequence-200990-        2976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobius Assembly Level 2 Acceptor Vector A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2976)
  AUTHORS   Andreou AI, Nakayama N
  TITLE     Mobius Assembly: A versatile Golden-Gate framework towards universal
            DNA assembly.
  JOURNAL   PLoS One. 2018 Jan 2;13(1):e0189892. doi: 
            10.1371/journal.pone.0189892. eCollection 2018.
  PUBMED    29293531
REFERENCE   2  (bases 1 to 2976)
  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: "PLoS One.";
            date: "2018-01-2"; volume: "13(1):e0189892. doi"; pages: " 
            10.1371/journal.pone.0189892. eCollection 2018"
COMMENT     Sequence Label: pMA_Level 2A
FEATURES             Location/Qualifiers
     source          1..2976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..65
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(69..112)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        143..177
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             186..197
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      928..999
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1015..1042
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1064..1121
                     /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(1145..1162)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1316..1904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1396..1415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2087..2181)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2202..2861)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2776..2795
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
       61 gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc
      121 ggtctcagga gatatgcagg tgtttacggc tagctcagtc ctaggtatag tgctagctac
      181 tagagaaaga ggagaaatac taaatgcgta aaggcgaaga actgttcacg ggcgtagttc
      241 cgattctggt cgagctggac ggcgatgtga acggtcataa gtttagcgtt cgcggtgaag
      301 gtgagggcga cgcgaccaac ggcaaactga ccctgaagtt catctgcacc accggtaaac
      361 tgccggtgcc ttggccgacc ttggtgacga cgttgacgta tggcgtgcag tgttttgcgc
      421 gttatccgga ccacatgaaa caacacgatt tcttcaaatc tgcgatgccg gagggttacg
      481 tccaggagcg taccatttcc ttcaaggatg atggctacta caaaactcgc gcagaggtta
      541 agtttgaagg tgacacgctg gtcaatcgta tcgaattgaa gggtatcgac tttaaagagg
      601 atggtaacat tctgggccat aaactggagt ataacttcaa cagccataat gtttacatta
      661 cggcagacaa gcaaaagaac ggcatcaagg ccaatttcaa gattcgccac aatgttgagg
      721 acggtagcgt ccaactggcc gaccattacc agcagaacac cccaattggt gacggtccgg
      781 ttttgctgcc ggataatcac tatctgagca cccaaagcgt gctgagcaaa gatccgaacg
      841 aaaaacgtga tcacatggtc ctgctggaat ttgtgaccgc tgcgggcatc acccacggta
      901 tggacgagct gtataagtaa tgagcttcaa ataaaacgaa aggctcagtc gaaagactgg
      961 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     1021 ccttcgggtg ggcctttctg cgcacctgca tatacccctg cagtccggca aaaaagggca
     1081 aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc gttatgcagg
     1141 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1201 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1261 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1321 acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1741 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2041 tctaaagtat atatgagtaa acttggtctg acagctcgag gcttggattc tcaccaataa
     2101 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     2161 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
     2221 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     2281 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     2341 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     2401 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     2461 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     2521 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     2581 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     2641 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     2701 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     2761 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     2821 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     2881 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     2941 ttggaacctc ttacgtgccc gatcaactcg agtgcc
//
