LOCUS       sequence-323966-        6836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal (His)6-TEV cleavage site fusion of human gene 
            or portion of gene in bacterial strains. pET based vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6836)
  TITLE     Davis Lab plasmids
REFERENCE   2  (bases 1 to 6836)
  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: thoc5.018.389
FEATURES             Location/Qualifiers
     source          1..6836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(569..1360,1361..1429))
                     /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     1192..1211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1430..1534)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1602..1620
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1647..1652,1653..1669,1670..1675)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(1850..1897)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1893..1911
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             2353..2364
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(3107..3127)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3140..3157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3167..3189
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3204..3228
                     /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(3228..3247)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3229..3247)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3331..3350)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3556..3633
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3634..4716
                     /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     complement(3653..3672)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             6028..6219
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6213..6235)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6321..6460
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6376..6395
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6475..6492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6646..6836,1..398))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6726..6745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
       61 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      241 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
      421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
      721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
      841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
      901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     1201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1561 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1621 cctttcgtct tcaagaattc tcatgtttga cagcttatca tcgataagct ttaatgcggt
     1681 agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg
     1741 ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt ggttatgccg
     1801 gtactgccgg gcctcttgcg ggatatccgg atatagttcc tcctttcagc aaaaaacccc
     1861 tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag
     1921 ccaactcagc ttcctttcgg gctttgttag cagccggatc ctcgagtgcg gccgcaagct
     1981 tcgtcatcac tagatggggg tgatcagctc catggcagtt gtcactttgg cttttactgt
     2041 catgatgttg aggttcatga ggtagtagaa agtcaggtga agcacactgt catctttgca
     2101 cttcaggtcg agcatgacag acagtgggtg cctcttcagc atctccttgc gtttgtcgtc
     2161 caactgaacc cccagtgtgg gtctccggcg cttcgtagtc tgctcctcct cggcatctga
     2221 gtcactctcg tcatcttggg agtcctctgg aggtttgaac agagccttgg cttcatccac
     2281 actgccttcg attgccacag ataacgtctt atcacaggcc tgcccatacg cagtggcctg
     2341 aacaaagagg acatagaggg gaggcggcag gtgtctggct gtctcatact gcttgtgagc
     2401 ctggtcgaat ggcataaaca ggtactcctg caccggaagg gaagcctgca tgatgctgtt
     2461 gaggcggggc tggaggctgc tcaggtactc cttcttcacc tcaatctcct tgagaatctt
     2521 ctccttgtta gataggcact ctcggtactt ctctgccagc cttttccgct gctccagctc
     2581 ccagtccaga cgtgccagtg tttgctggtg agggtctccc atggtgactt cggccttgct
     2641 gatatctggt ggagcctcct tataaaactc ctctaaactg accagatcaa tttcttcatg
     2701 ctttgactta aactccaaac atttggtgat ctccttctgt aggtgcatca cctcatacaa
     2761 caggttctgg agctgcagat gataggcatc tactttctgc ttagcctcgt gggtctgatc
     2821 tcttcctttc ttcaacctga tgtgggctaa tcggttaagc ttctttagag tcatgaaatg
     2881 cacacagctc tggatcctcc gttcttctat ttctattgcc acatccttgc cacccctgct
     2941 cttcaggtct tggatctcag ccatcagcct ctgtagctcc tggcaggtgt acttgtataa
     3001 ctcatagtct ctgccagggt cccgcagatc cacctcggcc tcctcactgt agtatttacc
     3061 ttcctgctcg gtgtcagatc gattccgctt tccttcagct ggggctgccc tggaaataca
     3121 agttttctct gccgctgctg tgatgatgat gatgatgcat ggtatatctc cttcttaaag
     3181 ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt
     3241 cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc gtggccggca
     3301 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     3361 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
     3421 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
     3481 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
     3541 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
     3601 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     3661 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     3721 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     3781 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     3841 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     3901 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     3961 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     4021 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     4081 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     4141 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     4201 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     4261 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     4321 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     4381 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     4441 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     4501 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     4561 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     4621 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     4681 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     4741 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
     4801 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     4861 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     4921 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     4981 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     5041 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
     5101 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     5161 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
     5221 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
     5281 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg
     5341 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     5401 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga
     5461 actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc
     5521 agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg
     5581 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
     5641 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
     5701 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg
     5761 ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca
     5821 cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc
     5881 cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca
     5941 tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac
     6001 agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg
     6061 gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg
     6121 gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc
     6181 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6241 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6301 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     6361 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat tgcggtgtga
     6421 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     6481 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6541 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgcttt
//
