LOCUS       sequence-466059-        5524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for 10x-phi29 DNA polymerase (PovSak): 
            PT7-lacO-g10leader-10xHis-p2 povsak-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: pTE5444
FEATURES             Location/Qualifiers
     source          1..5524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     161..180
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(200..277)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     483..502
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     586..605
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        586..604
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    605..629
                     /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             644..666
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             679..708
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             721..732
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      2505..2562
                     /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."
     primer_bind     complement(2586..2603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2757..3345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2837..2856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3528..3622)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3643..4302)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4217..4236
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4464..4482
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4486..4529)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4532..4553
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
       61 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
      121 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
      181 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
      241 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
      301 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
      361 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
      421 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
      481 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
      541 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
      601 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
      661 aggagatata ccatgggcca tcatcatcat catcatcatc atcatcacag cagcggccat
      721 atcgaaggtc gtcatgggtc tcgaatgccg cgtaaacgtt acagctgcga ttttgaaacg
      781 acgacgaaag ttgaagattg ccgtgtctgg gcctatggtt atatgaacat cgaagaccat
      841 tcagaatata aaattggcaa ctcgctggat gaatttatgg cgtgggcgct gaaagttcag
      901 gccgacctgt acttccacaa tctgaaattt gatggtgcgt tcattatcaa ctggctggaa
      961 cgtaatggct ttaaatggag cgccgatggt ctgccgaaca cctataatac gattatctct
     1021 cgtaccggcc aatggtatat gattgatatc tgcctgggct acaaaggtaa acgcaaaatt
     1081 cataccgtga tctatgacag cctgaaaaaa ctgccgtttc cggtgaagaa aattgcgaaa
     1141 gatttcaaac tgaccgtcct gaaaggcgat attgactatc acaaagaacg tccggttggt
     1201 tacaaaatca cgccggaaga atatgcgtac attaaaaacg atatccagat tatcgcagaa
     1261 gctctgctga ttcagtttaa acaaggcctg gatcgcatga ccgccggcag tgactccctg
     1321 aaagatttca aagatatcat caccacgaaa aaatttaaga aagtgttccc gaccctgagc
     1381 ctgggtctgg ataaaaaagt tcgttatgca taccgcggcg gttttacgtg gctgaacgac
     1441 cgtttcaaag aaaaagaaat tggcgagggt atggtctttg atgtgaatag tctgtatccg
     1501 gctcagatgt actcccgcct gctgccgtat ggcgaaccga tcgttttcga gggtaaatat
     1561 gtctgggatg aagactaccc gctgcatatt cagcacatcc gttgtgaatt tgaactgaaa
     1621 gaaggctata ttccgaccat tcaaatcaaa cgtagccgct tctataaggg taacgaatac
     1681 ctgaaaagct ctggcggtga aatcgcagac ctgtggctga gtaacgtcga tctggaactg
     1741 atgaaagaac attacgatct gtacaacgtt gaatacatct ccggcctgaa atttaaagcc
     1801 accacgggtc tgtttaaaga tttcattgac aaatggacct acatcaaaac cacgtctgaa
     1861 ggtgcaatca aacagctggc taaactgatg ctgaacagcc tgtatggcaa atttgcatct
     1921 aatccggatg ttaccggtaa agtcccgtac ctgaaagaaa atggcgctct gggttttcgc
     1981 ctgggcgaag aagaaaccaa agatccggtg tatacgccga tgggtgtttt cattaccgcg
     2041 tgggcacgtt acaccaccat caccgccgca caagcgtgct atgaccgcat tatctactgt
     2101 gataccgact caattcatct gaccggcacg gaaatcccgg atgtgattaa agatatcgtt
     2161 gacccgaaaa aactgggtta ttgggcacac gaatcgacct ttaaacgtgc taaatacctg
     2221 cgccagaaaa cgtacatcca agacatctac atgaaagaag tcgatggcaa actggtggaa
     2281 ggttcaccgg atgactatac cgacattaaa ttttcggtga aatgcgccgg catgaccgat
     2341 aaaattaaga aagaagtgac gttcgaaaat ttcaaagtgg gtttcagtcg caaaatgacc
     2401 ccgaaaccgg tccaagttcc gggcggcgtt gtgctggtcg ataacacctt cacgatcaaa
     2461 taagctttga gaccactcat cgcgactagt agcggccgct gcagtccggc aaaaaagggc
     2521 aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg cgttatgcag
     2581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2761 cacaggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3181 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3481 atctaaagta tatatgagta aacttggtct gacagctcga ggcttggatt ctcaccaata
     3541 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     3601 actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca
     3661 ctcgtcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     3721 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     3781 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     3841 ggtgaaactc acccagggat tggctgatac gaaaaacata ttctcaataa accctttagg
     3901 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     3961 ccggaaatcg tcgtggtatt cactccagag tgatgaaaac gtttcagttt gctcatggaa
     4021 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     4081 acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     4141 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     4201 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     4261 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     4321 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     4381 gttggaacct cttacgtgcc cgatcaactc gagtgccacc tgacgtctaa gaaaccatta
     4441 ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
     4501 aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta gagcgatgag
     4561 tctgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     4621 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaactg tcgtgccagc
     4681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     4741 gtttttcttt tcaccagtga gactggcaac agctgattgc ccttcaccgc ctggccctga
     4801 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     4861 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4921 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4981 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     5041 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     5101 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     5161 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     5221 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     5281 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     5341 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     5401 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     5461 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     5521 gtgg
//
