LOCUS       sequence-361468-        3449 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Human Mitochondrial SSB in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3449)
  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 3449)
  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..3449
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      1..87
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      179..206
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(301..317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(301..317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(314..336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    325..341
                     /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(349..355,356..373,374..379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    394..415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(532..549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(703..1291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(783..802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1462..2253,2254..2322))
                     /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     2085..2104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2638..3351)
                     /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             3440..3448
                     /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)"
ORIGIN
        1 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
       61 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
      121 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
      181 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctctttttgt ttatttttct
      241 aaatacattc aaatatgtat ccgctcatga gacaatagat ctaagcttgg cgtaatcatg
      301 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      361 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      421 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     2101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2341 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     2401 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     2461 ctccccgcgc ggacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2521 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2581 cgccatagac gtctttgaat tgttatcagc tatgcgccga ccagaacacc ttgccgatca
     2641 gccaaacgtc tcttcaggcc actgactagc gataactttc cccacaacgg aacaactctc
     2701 attgcatggg atcattgggt actgtgggtt tagtggttgt aaaaacacct gaccgctatc
     2761 cctgatcagt ttcttgaagg taaactcatc acccccaagt ctggctatgc agaaatcacc
     2821 tggctcaaca gcctgctcag ggtcaacgag aattaacatt ccgtcaggaa agcttggctt
     2881 ggagcctgtt ggtgcggtca tggaattacc ttcaacctca agccagaatg cagaatcact
     2941 ggcttttttg gttgtgctta cccatctctc cgcatcacct ttggtaaagg ttctaagctc
     3001 aggtgagaac atccctgcct gaacatgaga aaaaacaggg tactcatact cacttctaag
     3061 tgacggctgc atactaaccg cttcatacat ctcgtagatt tctctggcga ttgaagggct
     3121 aaattcttca acgctaactt tgagaatttt tgtaagcaat gcggcgttat aagcatttaa
     3181 tgcattgatg ccattaaata aagcaccaac gcctgactgc cccatcccca tcttgtctgc
     3241 gacagattcc tgggataagc caagttcatt tttctttttt tcataaattg ctttaaggcg
     3301 acgtgcgtcc tcaagctgct cttgtgttaa tggtttcttt tttgtgctca tacgttaaat
     3361 ctatcaccgc aagggataaa tatctaacac cgtgcgtgtt gactatttta cctctggcgg
     3421 tgataatggt tgcatgtact aaggaggtt
//
