LOCUS       Exported                6116 bp ds-DNA     circular SYN 24-FEB-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  AUTHORS   Haipei Liu
  TITLE     Direct Submission
  JOURNAL   Exported Feb 24, 2022 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6116
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      8..463
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(556..1371)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1493..2081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2511..2702)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3511..4593)
                     /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(4594..4671)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4980..4998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4999..5023
                     /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             5054..5059
     misc_feature    5068..5070
                     /label=M
     CDS             5071..5115
                     /codon_start=1
                     /label=Fg
                     /translation="NEEGFFSARGHRPLD"
     CDS             5140..5379
                     /codon_start=1
                     /label=FIVAR
                     /translation="DKTNLGELINQGKSLLDESVEGFNVGEYHKGAKDGLTVEINKAEE
                     VFNKEDATEEEINLAKESLEGAIARFNSLLIEEST"
     CDS             5404..5568
                     /codon_start=1
                     /label=ELP_MV7E2
                     /translation="VGVPGMGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG
                     EGVPGEGVPG"
     CDS             5569..5733
                     /codon_start=1
                     /label=ELP_MV7E2
                     /translation="VGVPGMGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG
                     EGVPGEGVPG"
     CDS             5734..5898
                     /codon_start=1
                     /label=ELP_MV7E2
                     /translation="VGVPGMGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG
                     EGVPGEGVPG"
     CDS             5908..5925
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5932..5964
                     /codon_start=1
                     /label=ybbR Tag
                     /translation="DSLEFIASKLA"
     terminator      6040..6087
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
       61 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      121 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      181 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
      241 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      301 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      361 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      421 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg
      481 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      541 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
      601 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
      661 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
      721 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
      781 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
      841 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
      901 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
      961 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     1021 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     1081 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1141 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1201 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     1261 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1321 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     1381 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     1441 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1501 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1561 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1621 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1681 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1741 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1801 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1861 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1921 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1981 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2041 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2101 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2161 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2221 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2281 ttctccttac gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat
     2341 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     2401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2521 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     2581 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     2641 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     2701 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     2761 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     2821 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     2881 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     2941 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3001 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3061 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3121 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     3181 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     3241 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     3301 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     3361 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     3421 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3481 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4261 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4561 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     4621 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     4681 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4741 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     4801 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     4861 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     4921 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     4981 aatacgactc actatagggg aattgtgagc ggataacaat tcccctgtag aaataatttt
     5041 gtttaacttt aagaaggaga tatacatatg aatgaagaag gcttttttag cgcacgtggt
     5101 catcgtccgc tggatggctc cggttctggt tccggctctg acaagacgaa cctgggtgaa
     5161 ctgatcaacc agggcaaatc tctgctggat gaatctgtag aaggtttcaa cgttggtgaa
     5221 taccacaagg gtgcaaaaga tggtctgact gttgaaatca acaaagcgga agaagtgttc
     5281 aacaaagaag acgccaccga agaggagatc aacctggcaa aagagagcct ggaaggtgct
     5341 atcgctcgtt tcaactctct gctgatcgaa gaatctaccg gctccggttc tggttcccac
     5401 ggcgtgggtg ttccaggtat gggtgtgcct ggtgttggtg ttcctggcgt tggcgtgcca
     5461 ggtgtgggag taccaggcgt aggcgttccc ggtgtaggcg tgcctggcgt cggtgtaccc
     5521 ggtgtcggag tccctggtga aggtgtgcca ggcgaaggtg tgccgggcgt gggtgttcca
     5581 ggtatgggtg tgcctggtgt tggtgttcct ggcgttggcg tgccaggtgt gggagtacca
     5641 ggcgtaggcg ttcccggtgt aggcgtgcct ggcgtcggtg tacccggtgt cggagtccct
     5701 ggtgaaggtg tgccaggcga aggtgtgccg ggcgtgggtg ttccaggtat gggtgtgcct
     5761 ggtgttggtg ttcctggcgt tggcgtgcca ggtgtgggag taccaggcgt aggcgttccc
     5821 ggtgtaggcg tgcctggcgt cggtgtaccc ggtgtcggag tccctggtga aggtgtgcca
     5881 ggcgaaggtg tgccgggctg gcgcggtcat catcatcacc atcatggtag cgactctctg
     5941 gaattcatcg cttctaaact ggcttaactc gagtaagatc cggctgctaa caaagcccga
     6001 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     6061 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggc
//
