LOCUS       sequence-366174-        3924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human Mitochondrial SSB in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3924)
  AUTHORS   Qian Y, Johnson KA
  TITLE     The human mitochondrial single-stranded DNA-binding protein displays
            distinct kinetics and thermodynamics of DNA binding and exchange.
  JOURNAL   J Biol Chem. 2017 Aug 4;292(31):13068-13084. doi: 
            10.1074/jbc.M117.791392. Epub 2017 Jun 14.
  PUBMED    28615444
REFERENCE   2  (bases 1 to 3924)
  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"; doi: 
            "10.1074/jbc.M117.791392"; journalName: "J Biol Chem"; date: 
            "2017-08-4- 4"; volume: "292"; issue: "31"; pages: "13068-13084"
COMMENT     Sequence Label: pcI-ts,ind+-(mtSSB delta20)
FEATURES             Location/Qualifiers
     source          1..3924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(1177..1890)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             1979..1987
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      2465..2551
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2643..2670
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2764..2780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2764..2780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2777..2799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2788..2804
                     /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(2812..2818,2819..2836,2837..2842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2857..2878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2995..3012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3166..3754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3246..3265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tatgtaagca gacagtttta
      901 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
      961 cgtggctttg ttgaataaat cgaacttttg ctgagttgac tccccgcgcg gacattaatt
     1021 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1081 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccatagacg tctttgaatt
     1141 gttatcagct atgcgccgac cagaacacct tgccgatcag ccaaacgtct cttcaggcca
     1201 ctgactagcg ataactttcc ccacaacgga acaactctca ttgcatggga tcattgggta
     1261 ctgtgggttt agtggttgta aaaacacctg accgctatcc ctgatcagtt tcttgaaggt
     1321 aaactcatca cccccaagtc tggctatgca gaaatcacct ggctcaacag cctgctcagg
     1381 gtcaacgaga attaacattc cgtcaggaaa gcttggcttg gagcctgttg gtgcggtcat
     1441 ggaattacct tcaacctcaa gccagaatgc agaatcactg gcttttttgg ttgtgcttac
     1501 ccatctctcc gcatcacctt tggtaaaggt tctaagctca ggtgagaaca tccctgcctg
     1561 aacatgagaa aaaacagggt actcatactc acttctaagt gacggctgca tactaaccgc
     1621 ttcatacatc tcgtagattt ctctggcgat tgaagggcta aattcttcaa cgctaacttt
     1681 gagaattttt gtaagcaatg cggcgttata agcatttaat gcattgatgc cattaaataa
     1741 agcaccaacg cctgactgcc ccatccccat cttgtctgcg acagattcct gggataagcc
     1801 aagttcattt ttcttttttt cataaattgc tttaaggcga cgtgcgtcct caagctgctc
     1861 ttgtgttaat ggtttctttt ttgtgctcat acgttaaatc tatcaccgca agggataaat
     1921 atctaacacc gtgcgtgttg actattttac ctctggcggt gataatggtt gcatgtacta
     1981 aggaggttca tatgactacc agtttggttc ttgaaagatc cctgaatcgt gtgcacttac
     2041 ttgggcgagt gggtcaggac cctgtcttga gacaggtgga aggaaaaaat ccagtcacaa
     2101 tattttctct agcaactaat gagatgtggc gatcagggga tagtgaagtt taccaactgg
     2161 gtgatgtcag tcaaaagaca acatggcaca gaatatcagt attccggcca ggcctcagag
     2221 acgtggcata tcaatatgtg aaaaaggggt ctcgaattta tttggaaggg aaaatagact
     2281 atggtgaata catggataaa aataatgtga ggcgacaagc aacaacaatc atagctgata
     2341 atattatatt tctgagtgac cagacgaaag agaaggagta gaaaggatga ttcttctttg
     2401 gccatcattt ggtacagtct catttccaag tcatgatggc atgcagtagg gaactgccag
     2461 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     2521 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     2581 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     2641 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     2701 tctaaataca ttcaaatatg tatccgctca tgagacaata gatctaagct tggcgtaatc
     2761 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2821 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2881 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2941 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3001 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3061 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3901 atatgagtaa acttggtctg acag
//
