LOCUS       pET51b_ADPHexTl         6586 bp    DNA     circular SYN 27-FEB-2025
DEFINITION  New Assembly 6586bp ds-DNA circular SYN  14-AUG-2024.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6586)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 24, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             29..58
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     terminator      95..142
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      179..634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(join(749..1540,1541..1609))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1610..1702)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1783..2371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2557..2699
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2801..2992)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3564..4646)
                     /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"
     promoter        complement(4647..4724)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        5037..5055
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5056..5080
                     /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             5095..5117
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(5133..6586,1..58)
                     /label=ADPHex T. lit.
     CDS             5133..5156
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5163..5177
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 agctccgggc ttctcctcaa tttccgctca tcaccaccat catcaccatc accaccacta
       61 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      121 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
      181 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      241 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      301 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      361 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      421 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      481 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      541 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      601 gcgaatttta acaaaatatt aacgtttaca atttctggcg gcacgatggc atgagattat
      661 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      721 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      781 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      841 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
      901 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      961 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1021 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1081 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1141 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1201 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1261 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1321 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1381 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1441 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1501 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1561 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1621 ttcaatcatg attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1681 tgtatttaga aaaataaaca aataggtcat gaccaaaatc ccttaacgtg agttttcgtt
     1741 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1801 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1861 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1921 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1981 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2041 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2101 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2161 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2221 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2281 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2341 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2401 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2461 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2521 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2581 gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat ctgctctgat
     2641 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     2701 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2761 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2821 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     2881 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     2941 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     3001 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     3061 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     3121 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     3181 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     3241 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     3301 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     3361 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     3421 gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac cggaaggagc
     3481 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     3541 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3601 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     3661 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     3721 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     3781 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     3841 gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     3901 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     3961 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4021 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4081 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4141 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4201 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4261 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4321 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4381 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     4441 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     4501 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     4561 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     4621 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     4681 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     4741 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     4801 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     4861 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     4921 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     4981 gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattaat
     5041 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     5101 taactttaag aaggagatat accatggcaa gctggagcca cccgcagttc gaaaagggtg
     5161 cagatgacga cgacaaggta ccgatgaagg aaagccttaa agataggata agattgtgga
     5221 aaaggctgta tgtaaacgcc tttgagaatg ccctaaacgc tatcccaaac gttaagggtg
     5281 ttcttctggc atataacacc aacattgatg ccataaaata cttagacaag gacgatctag
     5341 aaaagagagt aaccgaaata gggaaagaga aggtttttga aattattgaa aatccacctg
     5401 aaaagatctc ctccattgaa gagcttcttg gtggaatatt gaggagcata aaacttggca
     5461 aggctatgga gtggtttgta gagagtgagg aagtgaggag atacttaagg gaatggggat
     5521 gggatgagct gagaatagga gggcaggcag ggataatggc gaaccttctt gggggagtgt
     5581 atagaattcc aacgattgtt catgttcctc agaatccaaa gcttcaggcg gagttgtttg
     5641 tagatggtcc tatttatgtc cccgtctttg aaggaaataa gttgaagcta gttcacccta
     5701 aagatgccat tgcagaggaa gaagagttaa tccactacat atatgaattt cccagaggtt
     5761 ttcaggtttt tgatgttcaa gcaccaagag agaatagatt catagccaat gccgatgact
     5821 acaacgcgag ggtctacatg agaagggaat tcagagaagg ctttgaggaa attaccagaa
     5881 atgttgagct agcgataata agcgggctgc aggttttaaa ggaatactat cccgacggaa
     5941 caacatacag agatgttctt gatagagtcg aaagtcattt gaatatcctc aaccgctaca
     6001 acgtgaagag ccattttgaa tttgcatata ctgcaaacag aagggttagg gaggcacttg
     6061 tagagctttt accgaagttc acaagcgtcg gccttaatga agtagaactt gcctcaataa
     6121 tggagataat aggagacgag gagctggcaa aagaagttct ggagggacat atcttctcag
     6181 tcatagatgc gatgaatgtt ttaatggacg agacaggaat tgagagaatt cacttccaca
     6241 cctacggcta ctacctcgcc ctaactcaat acagaggaga agaggtaaga gatgctctgc
     6301 tctttgcatc cctcgctgca gctgccaaag ccatgaaagg aaaccttgaa agaatagagc
     6361 agattagaga tgccctaagt gtaccaacaa atgaaagggc aatagtgctt gaagaagaac
     6421 ttgaaaagga gttcacagaa tttgagaacg gccttattga tatggtggac agacaacttg
     6481 cttttgtgcc aacaaagatt gtagcatctc ccaagagcac cgttggaatt ggagatacca
     6541 tttcaagctc tgcttttgtc agtgaatttg gcatgaggaa aaggaa
//
