LOCUS       Exported                7510 bp ds-DNA     circular SYN 30-AUG-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7510)
  AUTHORS   Santana
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Aug 30, 2021 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7510
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     source          564..1400
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        65..83
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    84..108
                     /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             123..145
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             153..479
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             501..518
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             528..548
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    555..563
                     /label=linker
     misc_feature    564..656
                     /label=S1
                     /label=nonstandard type: SS
     misc_feature    657..749
                     /label=S2
                     /label=nonstandard type: SS
     misc_feature    750..842
                     /label=S3
                     /label=nonstandard type: SS
     misc_feature    843..935
                     /label=S4
                     /label=nonstandard type: SS
     misc_feature    936..1028
                     /label=S5
                     /label=nonstandard type: SS
     misc_feature    1029..1121
                     /label=S6
                     /label=nonstandard type: SS
     misc_feature    1122..1214
                     /label=S7
                     /label=nonstandard type: SS
     misc_feature    1215..1307
                     /label=S8
                     /label=nonstandard type: SS
     misc_feature    1308..1400
                     /label=S9
                     /label=nonstandard type: SS
     misc_feature    1401..1505
                     /label=P2
     misc_feature    1506..1613
                     /label=L2
     misc_feature    1614..1709
                     /label=S2
     misc_feature    1710..1811
                     /label=E1
     misc_feature    1812..1913
                     /label=E2
     CDS             1914..2321
                     /codon_start=1
                     /label=DYW
                     /translation="EPGCSWIEVKNKVHEFVAGDRSHPQSEEIYAKLEELSEKMKEAGY
                     VPDTSFVLHDVEEEEKEQMLSYHSEKLAIAFGLISTPPGTPIRIVKNLRVCGDCHTAIK
                     FISKIVGREIIVRDSNRFHHFKDGSCSCGDYW"
     CDS             2330..2369
                     /codon_start=1
                     /label=rpoA
                     /translation="EIEGTCITRAKSE"
     misc_feature    2351..2363
                     /label=rpoA binding site
                     /label=CLB19 binding site
     misc_feature    2364
                     /label=Edited C
     terminator      2382..2429
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2466..2921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2948..3052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3053..3913
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4084..4672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4858..5000
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5102..5293)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6102..7184)
                     /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(7185..7262)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct cgatcccgcg
       61 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
      121 attttgttta actttaagaa ggagatatac atatgagcga taaaattatt cacctgactg
      181 acgacagttt tgacacggat gtactcaaag cggacggggc gatcctcgtc gatttctggg
      241 cagagtggtg cggtccgtgc aaaatgatcg ccccgattct ggatgaaatc gctgacgaat
      301 atcagggcaa actgaccgtt gcaaaactga acatcgatca aaaccctggc actgcgccga
      361 aatatggcat ccgtggtatc ccgactctgc tgctgttcaa aaacggtgaa gtggcggcaa
      421 ccaaagtggg tgcactgtct aaaggtcagt tgaaagagtt cctcgacgct aacctggccg
      481 gttctggttc tggccatatg caccatcatc atcatcattc ttctggtgag aatctttatt
      541 ttcagggcgc catgggtaat tctgttgttt cttggaactc tatgatcact gcttatgcta
      601 gaaacggtag aattgaagag gctagaaagt tgtttgatga gatgcctgaa agagatgttg
      661 tttcttggaa tgctatgatc tctggatacg ctaaaaacgg tgagcttgat gaagctagaa
      721 agttgttcga tgagatgcca gagagggatg ttgtttcttg gacagctatg atctctggat
      781 atgctcaaaa tggtagaatt gatgaggcta gagaactttt cgataagatg cctgaaagaa
      841 acgttgtttc ttggaacgca atgatctctg gttacgctca gaacggtagg atcgatgagg
      901 ctagagaatt gttcgataag atgccagaga gatcagttgt ttcttggacc gcaatgatct
      961 ctggatacgc acagaatggt aggatagatg aagctagaga actttttgat aaaatgccag
     1021 agaggaatgt tgtttcttgg aatgcgatga tctctggtta tgcacagaac ggtaggatag
     1081 acgaagctag agaattgttt gataagatgc ctgaaagatc agttgtttct tggacggcaa
     1141 tgatctctgg ttatgctcag aatggtagga ttgacgaagc tagagaactt ttcgacaaaa
     1201 tgccagagcg tgacgttgtt tcttggaatg caatgatctc tggatatgca cagaacggta
     1261 gaatcgacga agctagagaa cttttcgaca agatgccaga gcgtgatgtt gtttcttgga
     1321 ctgctatgat ctctgcttat gctcaatgtg gacatttgga agaggctaga aagatttttg
     1381 atagaatgcc agagagagac gttgtttctt ggaatgcaat gatctctggt tacgctatgc
     1441 atggacatgg taaagaggct cttgaattgt ttgaagagat gcaacagtct ggtattaagc
     1501 cttctcatgt tactttcaca ggagttcttt ctgcttgttc tcatgctggt ttggttgatg
     1561 agggaagaca atacttcaac tctatgaaga aggattacgg tatcgaacca agagttgagc
     1621 attacggatg tatggttgat cttttgggaa gagctggtag acttgatgag gcttatgagt
     1681 ttattgagtc tatgcctatt gaaccaaatg ctgttgtttg gggtgctctt ttgggtgctt
     1741 gtaggattca tggaaacgtt gaattgggag agagagctgc tgagaaactt tttgaattgg
     1801 agccagaatc ttctggtaac tacgttcttt tgtctaacat ctatgcttct gctggaagat
     1861 gggatgatgt tgctaaggtt agaaaaatga tgaaggaaag aggtatcaag aaagagcctg
     1921 gatgttcttg gattgaagtt aagaataagg ttcatgagtt cgttgctgga gatagatcac
     1981 atccacaatc tgaagaaatc tatgctaaac ttgaagagtt gtctgaaaag atgaaagagg
     2041 ctggatatgt tcctgatact tcttttgttc ttcatgatgt tgaagaggaa gagaaagaac
     2101 agatgttgtc ttaccattct gagaaacttg ctatcgcttt cggtttgatt tctactcctc
     2161 caggaacacc aatcagaatc gttaaaaatc ttagagtttg tggagattgt catacagcta
     2221 ttaaattcat ctctaagatt gttggaagag aaattatcgt tagagattct aacagattcc
     2281 atcatttcaa ggatggttct tgttcttgtg gagattattg gtaactcgag aaatagaagg
     2341 aacatgtatt acacgtgcaa aatctgagag ctgagcaata actagcataa ccccttgggg
     2401 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
     2461 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2821 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2881 atttaacgcg aattttaaca aaatattaac gtttacaatt tcaggtggca cttttcgggg
     2941 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3001 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3061 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3121 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3181 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3241 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3301 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3361 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3421 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3481 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3541 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     3601 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3661 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3721 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3781 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3841 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3901 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3961 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4021 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4201 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4501 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4561 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4621 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4681 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4741 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4801 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     4861 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact
     4921 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     4981 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5041 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5101 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     5161 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     5221 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     5281 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     5341 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     5401 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     5461 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     5521 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     5581 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     5641 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     5701 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     5761 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     5821 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     5881 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     5941 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     6001 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     6061 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     6121 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6181 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     6241 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     6301 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     6361 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg
     6421 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     6481 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     6541 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     6601 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     6661 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     6721 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     6781 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     6841 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     6901 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     6961 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     7021 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     7081 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     7141 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     7201 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     7261 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     7321 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     7381 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     7441 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     7501 acctgtggcg
//
