LOCUS       sequence-199827-        3979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTPI1 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3979)
  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 3979)
  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-pTPI1 (01-11)
FEATURES             Location/Qualifiers
     source          1..3979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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)."
     primer_bind     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        97..115
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     97..114
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        616..1135
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     regulatory      1173..1182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1177..1186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1216..1235)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1217..1235)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1242..1259)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1242..1258)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1251..1273)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1399..1854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1486..1505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1696..1717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1932..2036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2037..2105,2106..2897)
                     /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     complement(2255..2274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3068..3656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3557..3576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3810..3827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3944..3965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagctattt aggtgacact atagaatact
      121 caagctatgc atccaacgcg ttgggagctc tcccatatgg tcgacctgca ggcggccgcg
      181 aattcactag tgattgggcg atgagtggtc tcgggagcta cgtatggtca tttcttcttc
      241 agattccctc atggagaaag tgcggcagat gtatatgaca gagtcgccag tttccaagag
      301 actttattca ggcacttcca tgataggcaa gagagaagac ccagagatgt tgttgtccta
      361 gttacacatg gtatttattc cagagtattc ctgatgaaat ggtttagatg gacatacgaa
      421 gagtttgaat cgtttaccaa tgttcctaac gggagcgtaa tggtgatgga actggacgaa
      481 tccatcaata gatacgtcct gaggaccgtg ctacccaaat ggactgattg tgagggagac
      541 ataactacat agtgtttaaa gattacggat atttaactta cttagaataa tgccattttt
      601 ttgagttata ataatcctac gttagtgtga gcgggattta aactgtgagg accttaatac
      661 attcagacac ttctgcggta tcaccctact tattcccttc gagattatat ctaggaaccc
      721 atcaggttgg tggaagatta cccgttctaa gacttttcag cttcctctat tgatgttaca
      781 cctggacacc ccttttctgg catccagttt ttaatcttca gtggcatgtg agattctccg
      841 aaattaatta aagcaatcac acaattctct cggataccac ctcggttgaa actgacaggt
      901 ggtttgttac gcatgctaat gcaaaggagc ctatatacct ttggctcggc tgctgtaaca
      961 gggaatataa agggcagcat aatttaggag tttagtgaac ttgcaacatt tactattttc
     1021 ccttcttacg taaatatttt tctttttaat tctaaatcaa tctttttcaa ttttttgttt
     1081 gtattctttt cttgcttaaa tctataacta caaaaaacac atacataaac taaaaccatc
     1141 gagaccactc atcgcccaat cgaattcccg cggccgccat ggcggccggg agcatgcgac
     1201 gtcgggccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     1261 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1321 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1381 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1441 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1501 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1561 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1621 atggttcacg tagtgggcca gcgccctgat agacggtttt tcgccctttg acgttggagt
     1681 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1741 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1801 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     1861 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatcaggt ggcacttttc
     1921 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1981 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2041 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2101 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2161 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2221 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2281 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2341 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2401 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2461 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2521 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2581 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2641 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2701 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtcgcgcg
     2761 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2821 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2881 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2941 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3001 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3061 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3121 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3181 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3241 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3301 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3361 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3421 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3481 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3541 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3601 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3661 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3721 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3781 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3841 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3901 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     3961 ctcattaggc accccaggc
//
