LOCUS       sequence-405671-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SSB1 protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  AUTHORS   Johnson LC, Singh A, Patel SS
  TITLE     The N-terminal domain of human mitochondrial helicase Twinkle has 
            DNA-binding activity crucial for supporting processive DNA synthesis
            by polymerase gamma.
  JOURNAL   J Biol Chem. 2023 Jan;299(1):102797. doi: 10.1016/j.jbc.2022.102797.
            Epub 2022 Dec 14.
  PUBMED    36528058
REFERENCE   2  (bases 1 to 6065)
  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: "J Biol 
            Chem."; date: "2023-01"; volume: "299(1):102797. doi"; pages: " 
            10.1016/j.jbc.2022.102797. Epub 2022 Dec 14"
COMMENT     Sequence Label: mt SSB
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     701..718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    733..872
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(798..817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     958..980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(974..1165)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2477..3559)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3521..3540
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3560..3637)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3843..3862
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3946..3965
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3946..3964
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3965..3989
                     /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)."
     RBS             4004..4026
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4511..4529)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4525..4572
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4747..4752,4753..4769,4770..4775))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4802..4820)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4888..4992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4993..5061,5062..5853)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5211..5230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6024..6065,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      781 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      841 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      901 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      961 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     1021 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     1081 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     1141 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     1201 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     1261 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     1321 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     1381 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     1441 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     1501 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     1561 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     1621 tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga
     1681 tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca
     1741 agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc
     1801 cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag
     1861 gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata
     1921 aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga
     1981 tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc
     2041 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg
     2101 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc
     2161 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     2221 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     2281 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     2341 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     2401 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     2461 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2521 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     2581 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     2641 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     2701 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     2761 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     2821 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     2881 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     2941 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     3001 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     3061 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     3121 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     3181 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     3241 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     3301 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     3361 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     3421 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     3481 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     3541 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     3601 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     3661 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     3721 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     3781 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     3841 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     3901 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     3961 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     4021 aggagatata catatggaat ccgaaacaac tactagtttg gttcttgaaa gatccctgaa
     4081 tcgtgtgcac ttacttgggc gagtgggtca ggaccctgtc ttgagacagg tggaaggaaa
     4141 aaatccagtc acaatatttt ctctagcaac taatgagatg tggcgatcag gggatagtga
     4201 agtttaccaa ctgggtgatg tcagtcaaaa gacaacatgg cacagaatat cagtattccg
     4261 gccaggcctc agagacgtgg catatcaata tgtgaaaaag gggtctcgaa tttatttgga
     4321 agggaaaata gactatggtg aatacatgga taaaaataat gtgaggcgac aagcaacaac
     4381 aatcatagct gataatatta tatttctgag tgaccagacg aaagagaagg agtagaaagg
     4441 atgattcttc tttggccatc ggatccggct gctaacaaag cccgaaagga agctgagttg
     4501 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4561 aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta
     4621 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
     4681 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
     4741 taccgcatta aagcttatcg atgataagct gtcaaacatg agaattcttg aagacgaaag
     4801 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4981 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5041 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5101 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5161 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5221 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5281 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5341 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5401 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5461 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5521 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5581 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5641 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5701 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5761 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5821 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5881 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5941 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6001 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6061 caaac
//
