LOCUS       sequence-199830-        3620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTEF2 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3620)
  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 3620)
  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-pTEF2 (01-11)
FEATURES             Location/Qualifiers
     source          1..3620
                     /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"
     regulatory      814..823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      818..827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(857..876)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(858..876)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(883..900)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(883..899)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(892..914)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1040..1495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1127..1146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1337..1358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1573..1677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1678..1746,1747..2538)
                     /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(1896..1915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2709..3297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3198..3217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3451..3468
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3585..3606
                     /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 tcgggagggg ccgtatactt acatatagta
      241 gatgtcaagc gtaggcgctt cccctgccgg ctgtgagggc gccataacca aggtatctat
      301 agaccgccaa tcagcaaact acctccgtac attcatgttg cacccacaca tttatacacc
      361 cagaccgcga caaattaccc ataaggttgt ttgtgacggc gtcgtacaag agaacgtggg
      421 aactttttag gctcaccaaa aaagaaagaa aaaatacgag ttgctgacag aagcctcaag
      481 aaaaaaaaaa ttcttcttcg actatgctgg aggcagagat gatcgagccg gtagttaact
      541 atatatagct aaattggttc catcaccttc ttttctggtg tcgctccttc tagtgctatt
      601 tctggctttt cctatttttt tttttccatt tttctttctc tctttctaat atataaattc
      661 tcttgcattt tctatttttc tctctatcta ttctacttgt ttattccctt caaggttttt
      721 ttttaaggag tacttgtttt tagaatatac ggtcaacgaa ctataattaa ctaaacccat
      781 cgagaccact catcgcccaa tcgaattccc gcggccgcca tggcggccgg gagcatgcga
      841 cgtcgggccc aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
      901 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
      961 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     1021 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1081 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1261 gatggttcac gtagtgggcc agcgccctga tagacggttt ttcgcccttt gacgttggag
     1321 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1381 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1441 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     1501 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt
     1561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcgcgc
     2401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2941 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3001 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3061 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3541 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     3601 actcattagg caccccaggc
//
