LOCUS       sequence-213442-        3253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyc1t in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3253)
  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 3253)
  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-Cyc1t (17-21)
FEATURES             Location/Qualifiers
     source          1..3253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..84
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     76..93
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     77..93
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     100..119
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        100..118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      149..158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      153..162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      200..391
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(494..512)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(495..512)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(530..546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(530..546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(543..565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    554..570
                     /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(578..584,585..602,603..608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    623..644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(761..778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(932..1520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1012..1031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1691..2482,2483..2551))
                     /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     2314..2333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2552..2656)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2734..3189)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2871..2892)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3083..3102
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
       61 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
      121 gaattgggcc cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgattggg
      181 cgatgagtgg tctcggctta tccgctctaa ccgaaaagga aggagttaga caacctgaag
      241 tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt
      301 tcaaattttt cttttttttc tgtacaaacg cgtgtacgca tgtaacatta tactgaaaac
      361 cttgcttgag aaggttttgg gacgctcgaa gcgctcgaga ccactcatcg cccaatcact
      421 agtgaattcg cggccgcctg caggtcgacc atatgggaga gctcccaacg cgttggatgc
      481 atagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg tcatagctgt
      541 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
      601 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      661 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      721 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      781 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      841 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      901 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      961 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1021 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1081 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1141 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1201 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1261 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1321 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1381 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1441 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1501 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1561 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1621 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1681 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1741 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1801 catctggccc cagtgctgca atgataccgc gcgacccacg ctcaccggct ccagatttat
     1861 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1921 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1981 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2041 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2101 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2161 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2221 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2281 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2341 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2401 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2461 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2521 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2581 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2641 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg
     2701 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa
     2761 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2821 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2881 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2941 agggcgctgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3001 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3061 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3121 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3181 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3241 ctcttcgcta tta
//
