LOCUS       sequence-466057-        5524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for 10x-phi29 DNA polymerase (PovSakOk): 
            PT7-lacO-g10leader-10xHis-p2 povsakok-T7 terminator, CamR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5524)
  AUTHORS   Barthel S, Hoffmann-Becking M, Karimov IG, Erb TJ
  TITLE     Genetically encoded control of in vitro transcription-translation 
            coupled DNA replication
  JOURNAL   bioRxiv 2025.07.08.663768; doi: 10.1101/2025.07.08.663768
REFERENCE   2  (bases 1 to 5524)
  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: "bioRxiv 
            2025.07.08.663768; doi: 10.1101/2025.07.08.663768"
COMMENT     Sequence Label: pTE5448
FEATURES             Location/Qualifiers
     source          1..5524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1100..1111)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(1124..1153)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1166..1188
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1203..1227
                     /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     complement(1227..1246)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1228..1246)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1330..1349)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1555..1632
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(1652..1671)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    2803..2824
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      2827..2870
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2874..2892)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             3054..3713
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3120..3139)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3734..3828
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4011..4599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4500..4519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4753..4770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4794..4851)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
ORIGIN
        1 tgtcaatgaa atctttaaac agacccgtgg tggctttaaa tttcaggccg gagatgtatt
       61 caacgttgta cagatcgtaa tgttctttca tcagttccag atcgacgtta ctcagccaca
      121 ggtctgcgat ttcaccgcca gagcttttca ggtattcgtt acccttatag aagcggctac
      181 gtttgatttg aatggtcgga atatagcctt ctttcagttc aaattcacaa cggatgtgct
      241 gaatatgcag cgggtagtct tcatcccaga catatttacc ctcgaaaacg atcggttcgc
      301 catacggcag caggcgggag tacatctgag ccggatacag actattcaca tcaaagacca
      361 taccctcgcc aatttctttt tctttgaaac ggtcgttcag ccacgtaaaa ccgccgcggt
      421 atgcataacg aactttttta tccagaccca ggctcagggt cgggaacact ttcttaaatt
      481 ttttcgtggt gatgatatct ttgaaatctt tcagggagtc actgccggcg gtcatgcgat
      541 ccaggccttg tttaaactga atcagcagag cttctgcgat aatctggata tcgtttttaa
      601 tgtacgcata ttcttccggc gtgattttgt aaccaaccgg acgttctttg tgatagtcaa
      661 tatcgccttt caggacggtc agtttgaaat ctttcgcaat tttcttcacc ggaaacggca
      721 gttttttcag gctgtcatag atcacggtat gaattttgcg tttacctttg tagcccaggc
      781 agatatcaat catataccat tggccggtac gagagataat cgtattatag gtgttcggca
      841 gaccatcggc gctccattta aagccattac gttccagcca gttgataatg aacgcaccat
      901 caaatttcag attgtggaag tacaggtcgg cctgaacttt cagcgcccac gccataaatt
      961 catccagcga gttgccaatt ttatattctg aatggtcttc gatgttcata taaccatagg
     1021 cccagacacg gcaatcttca actttcgtcg tcgtttcaaa atcgcagctg taacgtttac
     1081 gcggcattcg agacccatga cgaccttcga tatggccgct gctgtgatga tgatgatgat
     1141 gatgatgatg atggcccatg gtatatctcc ttcttaaagt taaacaaaat tatttctaga
     1201 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
     1261 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     1321 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     1381 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     1441 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     1501 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
     1561 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     1621 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     1681 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     1741 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     1801 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     1861 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     1921 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     1981 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     2041 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     2101 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     2161 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     2221 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     2281 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     2341 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     2401 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     2461 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     2521 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     2581 cagctgttgc cagtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     2641 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacagtt tcccgactgg
     2701 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     2761 tctcgaccga tgcccttgag agccttcaac ccagactcat cgctctagaa gcggccgcga
     2821 attccagaaa tcatccttag cgaaagctaa ggattttttt tatctgaaat tctgcctcgt
     2881 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2941 cactcgagtt gatcgggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     3001 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     3061 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     3121 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     3181 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     3241 ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac ggtgagctgg
     3301 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     3361 catcactctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     3421 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     3481 ttttcgtatc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     3541 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     3601 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     3661 gaatgcttaa tgaattacaa cagtactgcg acgagtggca gggcggggcg taatttgata
     3721 tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     3781 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcctcgag
     3841 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3901 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3961 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4021 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4081 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4141 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4201 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4261 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4321 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4381 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4441 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4501 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4561 tttttgtgat gctcgtcagg ggggcggagc ctgtggaaaa acgccagcaa cgcggccttt
     4621 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4681 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4741 acgaccgagc gcagcgagtc agtgagcgag gaagcctgca taacgcgaag taatcttttc
     4801 ggttttaaag aaaaagggca gggtggtgac accttgccct tttttgccgg actgcagcgg
     4861 ccgctactag tcgcgatgag tggtctcaaa gcttatttga tcgtgaaggt gttatcgacc
     4921 agcacaacgc cgcccggaac ttggaccggt ttcggggtca ttttgcgact gaaacccact
     4981 ttgaaatttt cgaacgtcac ttctttctta attttatcgg tcatgccggc gcatttcacc
     5041 gaaaatttaa tgtcggtata gtcatccggt gaaccttcca ccagtttgcc atcgacttct
     5101 ttcatgtaga tgtcttggat gtacgttttc tggcgcaggt atttagcacg tttaaaggtc
     5161 gattcgtgtg cccaataacc cagttttttc gggtcaacga tatctttaat cacatccggg
     5221 atttccgtgc cggtcagatg aattgagtcg gtatcacagt agataatgcg gtcatagcac
     5281 gcttgtgcgg cggtgatggt ggtgtaacgt gcccacgcgg taatgaaaac acccatcggc
     5341 gtatacaccg ggtttttggt ttcttcttcg cccaggcgaa aacccagagc gccattttct
     5401 ttcaggtacg ggactttacc ggtaacgttc ggattagatg caaatttgcc atacaggctg
     5461 ttcagcatca gtttagccag ctgtttgatt gcaccttcag acgtggtttt gatgtaggtc
     5521 catt
//
