LOCUS       sequence-466058-        5524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for 10xHis-phi29 DNA polymerase (wild-type): 
            PT7-lacO-g10leader-p2 wt-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: pTE5415
FEATURES             Location/Qualifiers
     source          1..5524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(171..182)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(195..224)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             237..259
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    274..298
                     /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(298..317)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(299..317)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(401..420)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        626..703
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(723..742)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    1874..1895
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1898..1941
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1945..1963)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             2125..2784
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2191..2210)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2805..2899
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3082..3670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3571..3590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3824..3841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3865..3922)
                     /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 gcctgaactt tcagcaccca cgccataaat tcatccagcg agttgccaat tttatattct
       61 gaatggtctt cgatgttcat ataaccatag gcccagacac ggcaatcttc aactttcgtc
      121 gtcgtttcaa aatcgcagct gtacatttta cgcggcattc gagacccatg acgaccttcg
      181 atatggccgc tgctgtgatg atgatgatga tgatgatgat gatggcccat ggtatatctc
      241 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
      301 tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc
      361 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
      421 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
      481 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
      541 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
      601 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
      661 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
      721 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
      781 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
      841 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
      901 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
      961 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     1021 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     1081 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     1141 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     1201 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     1261 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     1321 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     1381 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     1441 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     1501 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     1561 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     1621 actctctcag ggccaggcgg tgaagggcaa tcagctgttg ccagtctcac tggtgaaaag
     1681 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1741 aatgcagctg gcacgacagt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     1801 tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa
     1861 cccagactca tcgctctaga agcggccgcg aattccagaa atcatcctta gcgaaagcta
     1921 aggatttttt ttatctgaaa ttctgcctcg tgatacgcct atttttatag gttaatgtca
     1981 tgataataat ggtttcttag acgtcaggtg gcactcgagt tgatcgggca cgtaagaggt
     2041 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     2101 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     2161 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     2221 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     2281 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     2341 tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     2401 accgttttcc atgagcaaac tgaaacgttt tcatcactct ggagtgaata ccacgacgat
     2461 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     2521 tatttcccta aagggtttat tgagaatatg tttttcgtat cagccaatcc ctgggtgagt
     2581 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     2641 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     2701 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     2761 gacgagtggc agggcggggc gtaatttgat atcgagctcg cttggactcc tgttgataga
     2821 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc
     2881 gttttttatt ggtgagaatc caagcctcga gctgtcagac caagtttact catatatact
     2941 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3001 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3061 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3661 cctgtggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3781 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3841 ggaagcctgc ataacgcgaa gtaatctttt cggttttaaa gaaaaagggc agggtggtga
     3901 caccttgccc ttttttgccg gactgcagcg gccgctacta gtcgcgatga gtggtctcaa
     3961 agcttatttg atcgtgaagg tgtcatcgac cagcacaacg ccgcccggaa cttggaccgg
     4021 tttcggtttc attttgcgac tgaaacccac tttgaaattt tcgaacgtca cttctttctt
     4081 aattttatcg gtcatgccgg cgcatttcac cgaaaattta atgtcggtat agtcatccgg
     4141 tgaaccttcc accagtttgc catcgacttc tttcatgtag atgtcttgga tgtacgtttt
     4201 ctggcgcagg tatttagcac gtttaaaggt cgattcgtgt gcccaataac ccagtttttt
     4261 cgggtcaacg atatctttaa tcacatccgg gatttccgtg ccggtcagat gaattgagtc
     4321 ggtatcacag tagataatgc ggtcatagca cgcttgtgcg gcggtgatgg tggtgtaacg
     4381 tgcccacgcg gtaatgaaaa cacccatcgg cgtatacacc ggatctttgg tttcttcttc
     4441 gcccaggcga aaacccagag cgccattttc tttcaggtac gggactttac cggtaacatc
     4501 cggattagat gcaaatttgc catacaggct gttcagcatc agtttagcca gctgtttgat
     4561 tgcaccttca gacgtggttt tgatgtaggt ccatttgtca atgaaatctt taaacagacc
     4621 cgtggtggct ttaaatttca ggccggagat gtattcaacg ttgtacagat cgtaatgttc
     4681 tttcatcagt tccagatcga cgttactcag ccacaggtct gcgatttcac cgccagagct
     4741 tttcaggtat tcgttaccct tatagaagcg gctacgtttg atttgaatgg tcggaatata
     4801 gccttctttc agttcaaatt cacaacggat gtgctgaata tgcagcgggt agtcttcatc
     4861 ccagacatat ttaccctcga aaacgatcgg ttcgccatac ggcagcaggc gggagtacat
     4921 ctgagccgga tacagactat tcacatcaaa gaccataccc tcgccaattt ctttttcttt
     4981 gaaacggtcg ttcagccacg taaaaccgcc gcggtatgca taacgaactt ctttatccag
     5041 acccaggctc agggtcggga acactttctt aaattttttc gtggtgatga tatctttgaa
     5101 acctttcagg gagtcactgc cggcggtcat gcgatccagg ccttgtttaa actgaatcag
     5161 cagagcttct gcgataatct ggatatcgtt tttaatgtac gcatattctt ccggcgtgat
     5221 tttgtaacca accggacgtt ctttgtgata gtcaatatcg cctttcagga cggtcagttt
     5281 gaaatctttc gcaattttct tcaccggaaa cggcagtttt ttcaggctgt catagatcac
     5341 ggtatgaatt ttgcgtttac ctttgtagcc caggcagata tcaatcatat accattggcc
     5401 catacgagag ataatcgtat tataggtgtt cggcagacca tcggcgctcc atttaaagcc
     5461 attacgttcc agccagttga taatgaacgc accatcaaat ttcagattgt ggaagtacag
     5521 gtcg
//
