LOCUS       sequence-200992-        2980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobius Assembly Level 2 Acceptor Vector B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2980)
  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 2980)
  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 2B
FEATURES             Location/Qualifiers
     source          1..2980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(10..67)
                     /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."
     terminator      complement(89..116)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(132..203)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             934..945
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(954..988)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     terminator      1023..1066
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1070..1088)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1250..1909
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1316..1335)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1930..2024
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2207..2795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2696..2715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2949..2966
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgaagtaat cttttcggtt ttaaagaaaa agggcagggt ggtgacacct tgcccttttt
       61 tgccggactg caggggtata tgcaggtgcg cagaaaggcc cacccgaagg tgagccagtg
      121 tgactctagt agagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
      181 gactgagcct ttcgttttat ttgaagctca ttacttatac agctcgtcca taccgtgggt
      241 gatgcccgca gcggtcacaa attccagcag gaccatgtga tcacgttttt cgttcggatc
      301 tttgctcagc acgctttggg tgctcagata gtgattatcc ggcagcaaaa ccggaccgtc
      361 accaattggg gtgttctgct ggtaatggtc ggccagttgg acgctaccgt cctcaacatt
      421 gtggcgaatc ttgaaattgg ccttgatgcc gttcttttgc ttgtctgccg taatgtaaac
      481 attatggctg ttgaagttat actccagttt atggcccaga atgttaccat cctctttaaa
      541 gtcgataccc ttcaattcga tacgattgac cagcgtgtca ccttcaaact taacctctgc
      601 gcgagttttg tagtagccat catccttgaa ggaaatggta cgctcctgga cgtaaccctc
      661 cggcatcgca gatttgaaga aatcgtgttg tttcatgtgg tccggataac gcgcaaaaca
      721 ctgcacgcca tacgtcaacg tcgtcaccaa ggtcggccaa ggcaccggca gtttaccggt
      781 ggtgcagatg aacttcaggg tcagtttgcc gttggtcgcg tcgccctcac cttcaccgcg
      841 aacgctaaac ttatgaccgt tcacatcgcc gtccagctcg accagaatcg gaactacgcc
      901 cgtgaacagt tcttcgcctt tacgcattta gtatttctcc tctttctcta gtagctagca
      961 ctatacctag gactgagcta gccgtaaaca cctgcatatc tcctctgtga gaccgaattc
     1021 cagaaatcat ccttagcgaa agctaaggat tttttttatc tgaaattctg cctcgtgata
     1081 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     1141 cgagttgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1201 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1261 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1321 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1381 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1441 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     1501 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1561 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1621 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1681 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     1741 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     1801 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     1861 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga
     1921 gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1981 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc ctcgagctgt
     2041 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2101 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2161 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2221 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2281 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2341 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2401 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2461 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2521 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2581 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2641 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2701 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2761 tgtgatgctc gtcagggggg cggagcctgt ggaaaaacgc cagcaacgcg gcctttttac
     2821 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2881 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2941 ccgagcgcag cgagtcagtg agcgaggaag cctgcataac
//
