LOCUS       Exported                6905 bp ds-DNA     circular SYN 30-JUN-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 27, 2020 from SnapGene 5.1.2
            https://www.snapgene.com
COMMENT     >pET-15b  (5708 bp)
FEATURES             Location/Qualifiers
     source          1..6905
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
     promoter        10..38
                     /label=tet promoter
                     /note="tet promoter;E. coli promoter for tetracycline 
                     efflux protein gene"
     terminator      213..260
                     /label=T7 terminator
                     /note="T7 terminator;transcription terminator for 
                     bacteriophage T7 RNA polymerase"
     CDS             complement(330..1529)
                     /codon_start=1
                     /label=LanCL1
                     /note="LanCL1"
                     /translation="MAQRAFPNPYADYNKSLAEGYFDAAGRLTPEFSQRLTNKIRELLQ
                     QMERGLKSADPRDGTGYTGWAGIAVLYLHLYDVFGDPAYLQLAHGYVKQSLNCLTKRSI
                     TFLCGDAGPLAVAAVLYHKMNNEKQAEDCITRLIHLNKIDPHAPNEMLYGRIGYIYALL
                     FVNKNFGVEKIPQSHIQQICETILTSGENLARKRNFTAKSPLMYEWYQEYYVGAAAGLA
                     GIYYYLMQPSLQVSQGKLHSLVKPSVDYVCQLKFPSGNYPPCIGDNRDLLVHWCHGAPG
                     VIYMLIQAYKVFREEKYLCDAYQCADVIWQYGLLKKGYGLCHGSAGNAYAFLTLYNLTQ
                     DMKYLYRACKFAEWCLEYGEHGCRTPDTPFSLFEGMAGTIYFLADLLVPTKARFPAFEL
                     "
     misc_feature    561..566
                     /label=Cysteine Histidine Zinc Binding
                     /note="Cysteine Histidine Zinc Binding"
     misc_feature    702..704
                     /label=Cysteine Zinc Binding
                     /note="Cysteine Zinc Binding"
     primer_bind     complement(848..897)
                     /label=NL-pET15b-LanCL1-H219A-R
     primer_bind     859..904
                     /label=NL-pET15b-LanCL1-H219A-F
     misc_feature    873..875
                     /label=H219A
     CDS             complement(1533..1550)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin recognition and cleavage site
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(1560..1577)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis affinity tag
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(1587..1589)
                     /codon_start=1
                     /label=ATG
                     /note="ATG"
                     /translation="M"
     protein_bind    1633..1657
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator;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)."
     promoter        complement(1658..1676)
                     /label=T7 promoter
                     /note="T7 promoter;promoter for bacteriophage T7 RNA 
                     polymerase"
     promoter        1985..2062
                     /gene="lacI"
                     /label=lacI lacI promoter
                     /label=lacI
                     /note="lacI promoter"
     CDS             2063..3145
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /label=lac repressor
                     /note="lacI;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"
     CDS             4457..4648
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /label=Rop protein, which maintains plasmids at low copy 
                     number
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    4750..4892
                     /label=bom
                     /note="bom;basis of mobility region from pBR322"
     rep_origin      complement(5078..5666)
                     /direction=LEFT
                     /label=ori
                     /note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5837..6697)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /label=beta-lactamase
                     /note="AmpR;confers resistance to ampicillin, 
                     carbenicillin, and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6698..6802)
                     /gene="bla"
                     /label=bla AmpR promoter
                     /label=bla
                     /note="AmpR promoter"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctcgagt cagagttcaa atgcagggaa cctggctttt
      361 gtggggacta gcaggtcagc caggaaatat attgttccag ccattccttc aaagagagag
      421 aaaggggtgt ctggtgttct gcatccatgt tctccatact ctaagcacca ttcagcaaac
      481 ttacaggccc tatacaggta cttcatgtcc tgtgtgaggt tgtagagtgt caggaaggca
      541 taggcattcc ctgcagaacc gtggcacagc ccatatccct tcttcagcaa cccatattgc
      601 cagatcacat cagcacactg ataggcatca cagagatact tttcctctct gaatacctta
      661 taggcctgga tgagcatgta gattacccca ggggcgccat ggcaccaatg gacaagcaga
      721 tctcgattat cacctataca tggagggtaa ttgccagaag ggaatttcag ctggcagacg
      781 tagtctacac tgggcttgac caaactatgt aacttccctt ggctcacttg aaggctgggc
      841 tgcatcaggt agtaataaat tccagccagg ccggcagcag cccctacata atattcctgg
      901 taccattcat acatcagtgg agactttgcc gtgaagtttc tcttcctagc taggttttct
      961 ccagaggtta aaattgtttc acaaatctgc tgaatatggc tttgaggaat cttttccact
     1021 ccaaagttct tattgacaaa aagaagagca tagatgtagc ctattcgccc atagagcatt
     1081 tcatttggag catgaggatc aatcttattt aggtgaatta gccgtgtgat gcaatcttct
     1141 gcctgcttct cattgttcat cttgtgatat agcacagcgg ccactgccag ggggcctgca
     1201 tccccacaaa ggaaggtgat ggagcgcttg gttaagcagt tcagactttg ctttacatag
     1261 ccatgtgcta actgtaggta ggcagggtcc ccaaatacat cataaagatg taagtaaagc
     1321 acagcaatac ctgcccagcc agtgtaaccg gtgccatccc gagggtctgc tgatttcagg
     1381 cctctctcca tttgctgaag aagctcccga atcttattgg tcaagcgttg tgagaactca
     1441 ggagtcagcc tcccggcagc atcaaagtag ccttcggcca gggatttgtt ataatcagca
     1501 taaggattcg ggaaggccct ttgagccata tggctgccgc gcggcaccag gccgctgctg
     1561 tgatgatgat gatgatggct gctgcccatg gtatatctcc ttcttaaagt taaacaaaat
     1621 tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
     1681 cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     1741 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     1801 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     1861 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
     1921 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat
     1981 cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     2041 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     2101 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     2161 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     2221 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     2281 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     2341 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     2401 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     2461 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     2521 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     2581 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     2641 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     2701 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     2761 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     2821 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     2881 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     2941 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     3001 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     3061 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3121 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     3181 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     3241 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     3301 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     3361 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     3421 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     3481 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
     3541 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
     3601 ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca
     3661 gcctaacttc gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca
     3721 catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt
     3781 tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc
     3841 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
     3901 gcaaaccaac ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg
     3961 gcgcatctcg ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt
     4021 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
     4081 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
     4141 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
     4201 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     4261 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     4321 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     4381 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
     4441 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     4501 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     4561 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     4621 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4681 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4741 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     4801 tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt gaaataccgc
     4861 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     4921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5821 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5941 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6121 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     6181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6421 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     6481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6661 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6841 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     6901 aagaa
//
