LOCUS       Exported                5831 bp ds-DNA     circular SYN 03-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5831)
  AUTHORS   Lahcluster
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 3, 2021 from SnapGene Viewer 5.1.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5831
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             123..140
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    153..599
                     /label=LahT150
                     /note="LahT150"
     promoter        665..683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    684..708
                     /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)."
     CDS             817..861
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      913..960
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      997..1452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1567..2427)
                     /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(2428..2520)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2601..3189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3375..3517
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3619..3810)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4382..5464)
                     /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(5465..5542)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
      121 gccatcacca tcatcaccac agccaggatc cgagtaaaaa gcagatacag cctgtcacaa
      181 gagggcgagc caaggttcct gttatcatgc agatggaagc tctcgaatgt ggcgcggcat
      241 cacttgcaat ggtgcttgct tattataaaa aatgggtacc gctcgagcag gtcagagttg
      301 actgcggtgt atctcgtgac ggttccaatg cactgaatgt gcttaaggca gcacgaaact
      361 acggtcttga agcgaagggt tatcgttatg aaccggagaa gctgaagaag gaaggcacct
      421 tcccgtgtat tatacactgg aattttaatc attttgttgt tcttaaaggt ttcaagggga
      481 aatatgcata tataaacgat cctgcaaagg gtgatgttaa aataccgatg gaggagtttg
      541 atcgttcttt tacgggtata tgtctcattt tcaagcctac tgatagattt gaacagtaag
      601 cggccgcata atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc cgcataatcg
      661 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc atcttagtat
      721 attagttaag tataagaagg agatatacat atggcagatc tcaattggat atcggccggc
      781 cacgcgatcg ctgacgtcgg taccctcgag tctggtaaag aaaccgctgc tgcgaaattt
      841 gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc
      901 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      961 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     1021 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1081 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1141 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1201 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1261 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1321 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1381 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1441 cgtttacaat ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat
     1501 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1561 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1621 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1681 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1741 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1801 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1861 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1921 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1981 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2041 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2101 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2161 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2221 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2281 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2341 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2401 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt
     2461 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2521 taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3361 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3421 cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3481 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3541 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3601 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     3661 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     3721 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     3781 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     3841 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     3901 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     3961 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     4021 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4081 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     4141 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     4201 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4261 gatcatgcta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     4321 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     4381 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4441 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     4501 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     4561 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     4621 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     4681 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     4741 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     4801 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     4861 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     4921 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     4981 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     5041 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5101 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     5161 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     5221 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     5281 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     5341 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     5401 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     5461 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5521 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     5581 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     5641 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     5701 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     5761 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     5821 tagaggatcg a
//
