LOCUS       sequence-200993-        2980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobius Assembly Level 2 Acceptor Vector Gamma.
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 2-Gamma
FEATURES             Location/Qualifiers
     source          1..2980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(8..65)
                     /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(87..114)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(130..201)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             932..943
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(952..986)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     terminator      1021..1064
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1068..1086)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1248..1907
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1314..1333)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1928..2022
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2205..2793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2694..2713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2947..2964
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaagtaatct tttcggtttt aaagaaaaag ggcagggtgg tgacaccttg cccttttttg
       61 ccggactgca ggggtatatg caggtgcgca gaaaggccca cccgaaggtg agccagtgtg
      121 actctagtag agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
      181 ctgagccttt cgttttattt gaagctcatt acttatacag ctcgtccata ccgtgggtga
      241 tgcccgcagc ggtcacaaat tccagcagga ccatgtgatc acgtttttcg ttcggatctt
      301 tgctcagcac gctttgggtg ctcagatagt gattatccgg cagcaaaacc ggaccgtcac
      361 caattggggt gttctgctgg taatggtcgg ccagttggac gctaccgtcc tcaacattgt
      421 ggcgaatctt gaaattggcc ttgatgccgt tcttttgctt gtctgccgta atgtaaacat
      481 tatggctgtt gaagttatac tccagtttat ggcccagaat gttaccatcc tctttaaagt
      541 cgataccctt caattcgata cgattgacca gcgtgtcacc ttcaaactta acctctgcgc
      601 gagttttgta gtagccatca tccttgaagg aaatggtacg ctcctggacg taaccctccg
      661 gcatcgcaga tttgaagaaa tcgtgttgtt tcatgtggtc cggataacgc gcaaaacact
      721 gcacgccata cgtcaacgtc gtcaccaagg tcggccaagg caccggcagt ttaccggtgg
      781 tgcagatgaa cttcagggtc agtttgccgt tggtcgcgtc gccctcacct tcaccgcgaa
      841 cgctaaactt atgaccgttc acatcgccgt ccagctcgac cagaatcgga actacgcccg
      901 tgaacagttc ttcgccttta cgcatttagt atttctcctc tttctctagt agctagcact
      961 atacctagga ctgagctagc cgtaaacacc tgcatatctc ctgactgaga ccgaattcca
     1021 gaaatcatcc ttagcgaaag ctaaggattt tttttatctg aaattctgcc tcgtgatacg
     1081 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcactcg
     1141 agttgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     1201 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1261 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1321 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1381 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1441 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
     1501 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1561 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1621 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1681 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1741 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1801 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1861 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc
     1921 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     1981 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcct cgagctgtca
     2041 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2101 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2161 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2221 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2281 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2341 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2401 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2461 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2521 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2581 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2641 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2701 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2761 tgatgctcgt caggggggcg gagcctgtgg aaaaacgcca gcaacgcggc ctttttacgg
     2821 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2881 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2941 gagcgcagcg agtcagtgag cgaggaagcc tgcataacgc
//
