LOCUS       sequence-175058-        6796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET LIC cloning vector for Biotin-His6-tev-yORF-tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6796)
  TITLE     C-terminal fluorescent fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 6796)
  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"
COMMENT     Sequence Label: pET tdTomato LIC cloning vector (H6-tdTomato)
FEATURES             Location/Qualifiers
     source          1..6796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     705..724
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(744..821)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1027..1046
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1130..1149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1130..1148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1149..1173
                     /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             1188..1210
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1223..1267
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1268..1285
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1286..1306
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2789..2806
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2859..2877)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2873..2920
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2957..3412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3044..3063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3254..3275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3505..4320)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3505..4320)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4228..4247
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4312..4331
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4442..5030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4931..4950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5184..5201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5216..5355
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5281..5300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5441..5463
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5457..5648)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6457..6796,1..743))
                     /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"
ORIGIN
        1 tgatcgttgg caaccagcat cgcagtggga acgataccct cattcagcat ttgcatggtt
       61 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
      121 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
      181 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
      241 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
      301 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
      361 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
      421 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
      481 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
      541 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
      601 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
      661 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
      721 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
      781 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
      841 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
      901 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
      961 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     1021 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     1081 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     1141 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     1201 aagaaggaga tataccatgg gaggcctgaa cgatattttt gaagcgcaga aaattgaatg
     1261 gcatgaacac catcaccatc accatgaaaa cctgtacttc caatccaata ttggtagtgg
     1321 gagcaacggc agcagcggat ccgtgagtaa gggcgaggaa gtgatcaaag agttcatgcg
     1381 gtttaaggtg agaatggaag gaagcatgaa cggccacgag ttcgaaattg agggagaagg
     1441 agagggacgg ccctacgagg gcacccagac agccaagctg aaagtgacaa agggcgggcc
     1501 tctgccattc gcttgggaca tcctgagccc acagtttatg tacggctcca aggcctatgt
     1561 gaaacatcca gctgacattc ccgattataa gaaactgagc ttccccgagg ggtttaagtg
     1621 ggaaagagtg atgaacttcg aggacggagg cctggtgact gtgacccagg acagctccct
     1681 gcaggatggg accctgatct acaaggtgaa aatgagaggg acaaattttc cccctgatgg
     1741 acctgtgatg cagaagaaaa ctatgggatg ggaggcctcc accgaaaggc tgtatccacg
     1801 cgacggggtg ctgaaaggag aaatccacca ggctctgaag ctgaaagatg ggggacatta
     1861 cctggtggag ttcaagacaa tctacatggc caagaaacct gtgcagctgc caggctacta
     1921 ttacgtggac acaaaactgg atatcacttc acacaacgag gactacacta ttgtggagca
     1981 gtatgaacgg agcgagggga gacaccatct gttcctgggc catgggactg gaagtaccgg
     2041 ctcagggtct agtggaaccg cctcaagcga ggataacaat atggctgtga tcaaagagtt
     2101 catgaggttt aaggtgcgca tggagggcag catgaatggg cacgaatttg agattgaagg
     2161 agagggcgaa gggaggcctt acgagggcac acagactgcc aagctgaaag tgaccaaggg
     2221 aggaccactg cctttcgctt gggatatcct gtctcctcag tttatgtacg gaagtaaggc
     2281 ctatgtcaag catcccgctg acattcctga ttacaagaaa ctgtctttcc cagagggctt
     2341 taagtgggag agagtgatga attttgaaga tggaggcctg gtgaccgtga cacaggactc
     2401 ctctctgcag gatggcactc tgatctacaa agtcaaaatg cgcggcacca attttccacc
     2461 cgatgggccc gtgatgcaga agaaaacaat ggggtgggag gccagcactg aacggctgta
     2521 tcctagagac ggagtgctga agggcgaaat ccaccaggcc ctgaagctga aagacggcgg
     2581 ccactacctg gtggagttca aaaccatcta catggccaag aaaccagtgc agctgcccgg
     2641 ctattactat gtggacacca agctggatat cacatcccac aatgaagact acaccattgt
     2701 ggaacagtat gagaggtctg aaggacgcca ccatctgttt ctgtacggca tggatgagct
     2761 gtataagtaa taagcggccg cactcgagca ccaccaccac caccactgag atccggctgc
     2821 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     2881 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     2941 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3301 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3361 atttaacaaa aatttaacgc gaattttaac aaactagtaa cgtttacaat ttcaggtggc
     3421 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3481 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     3541 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3601 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3661 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3721 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     3781 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3841 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3901 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3961 aatgttttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     4021 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     4081 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     4141 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     4201 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     4261 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     4321 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc
     4381 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4441 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4501 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4561 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4621 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4681 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4741 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4801 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4861 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4921 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4981 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5041 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5101 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5161 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     5221 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     5281 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     5341 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5401 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5461 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     5521 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     5581 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     5641 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     5701 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     5761 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     5821 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     5881 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     5941 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     6001 cagcagtcgc ttcacgttcg ctcgcgtatc ggttattcat tctgctaacc agtaaggcaa
     6061 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     6121 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     6181 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     6241 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     6301 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     6361 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     6421 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6481 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6541 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     6601 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     6661 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     6721 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     6781 tgcgcccagc gccatc
//
