LOCUS       sequence-213437-        3202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAM33MTS  in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3202)
  AUTHORS   Perez-Gonzalez A, Kniewel R, Veldhuizen M, Verma HK, 
            Navarro-Rodriguez M, Rubio LM, Caro E
  TITLE     Adaptation of the GoldenBraid modular cloning system and creation of
            a toolkit for the expression of heterologous proteins in yeast 
            mitochondria.
  JOURNAL   BMC Biotechnol. 2017 Nov 13;17(1):80. doi: 
            10.1186/s12896-017-0393-y.
  PUBMED    29132331
REFERENCE   2  (bases 1 to 3202)
  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: "BMC 
            Biotechnol."; date: "2017-11-13"; volume: "17(1):80. doi"; pages: " 
            10.1186/s12896-017-0393-y"
COMMENT     Sequence Label: pUPD-MAM33MTS (12)
FEATURES             Location/Qualifiers
     source          1..3202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..96
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     88..105
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     89..105
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     112..131
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        112..130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      161..170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      165..174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(455..473)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(456..473)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(491..507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(491..507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(504..526)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    515..531
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(539..545,546..563,564..569))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    584..605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(722..739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(893..1481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(973..992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1652..2443,2444..2512))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2275..2294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2513..2617)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2695..3150)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2832..2853)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3044..3063
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
       61 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
      121 tcactatagg gcgaattggg cccgacgtcg catgctcccg gccgccatgg cggccgcggg
      181 aattcgattg ggcgatgagt ggtctcgcca tgttcttaag aagcgttaac cgtgccgtca
      241 ctagaagcat tttgactacg cccaagccag ccgtagtaaa atcatcgtgg agagttttta
      301 ctgttgctaa ctctaagaga tgtttcacac ccgctgcaat catgaggaac tcaatgcgag
      361 accactcatc gcccaatcac tagtgaattc gcggccgcct gcaggtcgac catatgggag
      421 agctcccaac gcgttggatg catagcttga gtattctata gtgtcaccta aatagcttgg
      481 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      541 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      601 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      661 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      721 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      781 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      841 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      901 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      961 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1021 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1081 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1141 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1201 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1261 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1321 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1381 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1441 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1501 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1561 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1621 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1681 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1741 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgcgacccac
     1801 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1861 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1921 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1981 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2041 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2101 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2161 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2221 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2281 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2341 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2401 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2461 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2521 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2581 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2641 ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     2701 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2761 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2821 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2881 aagggcgaaa aaccgtctat cagggcgctg gcccactacg tgaaccatca ccctaatcaa
     2941 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3001 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3061 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3121 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     3181 ggaagggcga tcggtgcggg cc
//
