LOCUS       Exported                5651 bp ds-DNA     circular SYN 03-MAR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5651)
  AUTHORS   LS fuer Angewandte Physik LMU - Lukas Milles
  TITLE     Direct Submission
  JOURNAL   Exported Aug 28, 2017 from SnapGene 4.0.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5651
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      9..464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(557..1372)
                     /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      1494..2082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2268..2410
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2512..2703)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3512..4594)
                     /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(4595..4672)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4957..4976
                     /label=T7 FW
     primer_bind     complement(4963..4985)
                     /label=rv from t7fw2
     promoter        4981..4985
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4986..5008
                     /label=T7FW#2
     primer_bind     4986..5008
                     /label=T7FW#2
     promoter        4986..4999
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5000..5024
                     /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)."
     primer_bind     complement(5037..5071)
                     /label=IV#440 Rev w/o N-tag pET28a
                     /note="BoxIV#440 Rev w/o N-tag pET28a"
     primer_bind     5037..5071
                     /label=IV#676 FW w/o N-tag pet28a
                     /note="BoxIV#676 FW w/o N-tag pet28a"
     primer_bind     complement(5040..5070)
                     /label=III#349 REV-w/o N-Tags MCS Met
                     /note="BoxIII#349 REV-w/o N-Tags MCS Met"
     primer_bind     complement(5042..5120)
                     /label=II#115 REV HIS
                     /note="BoxII#115 REV HIS"
     primer_bind     complement(5045..5104)
                     /label=REV ybbR
     primer_bind     complement(5054..5072)
                     /label=REV-w/o N-Tags MCS
     RBS             5055..5060
     CDS             5069..5071
                     /codon_start=1
                     /label=ATG
                     /translation="M"
     primer_bind     5072..5091
                     /label=FW ybbR
     CDS             5073..5104
                     /codon_start=1
                     /label=ybbR Tag
                     /translation="SLEFIASKLA"
     primer_bind     5103..5151
                     /label=fw ddFLN4 GSHIS GA
     primer_bind     complement(5103..5128)
                     /label=rv GS HIS
     CDS             5105..5122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     5122..5151
                     /label=fw ddFLN4 8GS
     CDS             5129..5431
                     /codon_start=1
                     /label=ddFLN4
                     /translation="ADPEKSYAEGPGLDGGESFQPSKFKIHAVDPDGVHRTDGGDGFVV
                     TIEGPAPVDPVMVDNGDGTYDVEFEPKEAGDYVINLTLDGDNVNGFPKTVTVKPAP"
     primer_bind     5129..5151
                     /label=fw ddFLN4
     primer_bind     complement(5149..5168)
                     /label=QC ddFLN4 C18S rv
     primer_bind     5169..5196
                     /label=QC ddFLN4 C18S
     CDS             5180..5182
                     /codon_start=1
                     /label=C18S
                     /translation="S"
     primer_bind     complement(5320..5341)
                     /label=rv ddFLN4add5G
     CDS             5339..5341
                     /codon_start=1
                     /label=Add 5G
                     /translation="P"
     primer_bind     5342..5365
                     /label=fw ddFLN4ad5G
     primer_bind     complement(5411..5455)
                     /label=rv ddFLN4 8GSXdoc
     primer_bind     complement(5411..5455)
                     /label=IV#682 rv ddFLN4 8GSXdoc
                     /note="BoxIV#682 rv ddFLN4 8GSXdoc"
     primer_bind     complement(5411..5444)
                     /label=rv ddFLN4 linkdoc1
     primer_bind     complement(5411..5434)
                     /label=rv ddFLN4 linker
     primer_bind     complement(5411..5434)
                     /label=IV#567 rv ddFLN4 linker
                     /note="BoxIV#567 rv ddFLN4 linker"
     primer_bind     complement(5411..5431)
                     /label=rv ddFLN4
     CDS             5432..5455
                     /codon_start=1
                     /label=8GS
                     /translation="GSGSGSGS"
     primer_bind     5432..5450
                     /label=fw 8GS to HIS
     primer_bind     complement(5437..5472)
                     /label=rv QC Fgbeta3 toC
     primer_bind     complement(5437..5455)
                     /label=rv Fgateb C
     primer_bind     complement(5437..5455)
                     /label=rv QC DK
     primer_bind     complement(5437..5455)
                     /label=rv QC K10
     primer_bind     complement(5437..5455)
                     /label=rv QC Fg alpha
     primer_bind     complement(5437..5455)
                     /label=rv 8GSandAT
     primer_bind     complement(5437..5455)
                     /label=IV#570 rv 8GS
                     /note="BoxIV#570 rv 8GS"
     primer_bind     complement(5437..5455)
                     /label=rv 8GS
     primer_bind     complement(5438..5455)
                     /label=rv QC nebpept toGS
     primer_bind     complement(5438..5455)
                     /label=rv 8GS dd4 ad SII
     CDS             5456..5500
                     /codon_start=1
                     /label=Fg beta
                     /translation="NEEGFFSARGHRPLD"
     primer_bind     5456..5479
                     /label=fw Fgb
     primer_bind     5471..5489
                     /label=fw QC FgB-F
     primer_bind     5477..5524
                     /label=fw QC Fgbeta3 toC
     primer_bind     5500..5523
                     /label=fw QC nebpept
     primer_bind     5501..5524
                     /label=fw QC DK
     primer_bind     5501..5524
                     /label=fw QC K10
     primer_bind     5501..5524
                     /label=fw QC Fg alpha
     primer_bind     5501..5523
                     /label=FW-w/o C-Tags MCS
     primer_bind     5501..5522
                     /label=fw STOP dd4 to sII
     primer_bind     5501..5521
                     /label=608 FW (LPTEGG) MCS GA
     primer_bind     complement(5501..5518)
                     /label=REV w/o C Tags
     primer_bind     complement(5558..5576)
                     /label=T7REV#2
     terminator      5576..5623
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5623..5646)
                     /label=T7 REV
ORIGIN
        1 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
       61 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      121 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      181 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      241 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      301 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      361 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      421 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcaggtg gcacttttcg
      481 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      541 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
      601 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
      661 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
      721 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
      781 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
      841 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
      901 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
      961 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     1021 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     1081 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     1141 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     1201 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     1261 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     1321 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     1381 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     1441 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1501 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1561 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1621 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1681 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1741 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1801 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1861 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1921 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1981 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2041 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2101 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2161 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2221 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     2281 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     2341 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     2401 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2461 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2521 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     2581 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     2641 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     2701 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     2761 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     2821 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     2881 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     2941 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     3001 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     3061 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     3121 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     3181 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     3241 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     3301 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     3361 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     3421 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     3481 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3541 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3601 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     3661 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     3721 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     3781 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     3841 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     3901 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     3961 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     4021 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     4081 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     4141 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     4201 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     4261 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     4321 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     4381 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     4441 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     4501 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     4561 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     4621 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     4681 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     4741 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     4801 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     4861 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     4921 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
     4981 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
     5041 tgtttaactt taagaaggag atatacatat ggactctctg gaattcatcg cttctaaact
     5101 ggctcaccac caccaccacc acggttctgc tgacccggaa aaatcttacg ctgaaggtcc
     5161 gggtctggac ggtggtgaaa gtttccagcc gtctaaattc aaaatccacg ctgttgaccc
     5221 ggacggtgtt caccgtaccg acggtggtga cggtttcgtt gttaccatcg aaggtccggc
     5281 tccggttgac ccggttatgg ttgacaacgg tgacggtacc tacgacgttg aattcgaacc
     5341 gaaagaagct ggtgactacg ttatcaacct gaccctggac ggtgacaacg ttaacggttt
     5401 cccgaaaacc gttaccgtta aaccggctcc gggctccggt tctggttccg gctctaatga
     5461 agagggcttt ttcagcgcac gtggtcatcg tccgctggat taactcgagt aagatccggc
     5521 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     5581 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     5641 atccggattg g
//
