LOCUS       Exported                5681 bp ds-DNA     circular SYN 28-JUL-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5681)
  AUTHORS   LS fuer Angewandte Physik LMU
  TITLE     Direct Submission
  JOURNAL   Exported Aug 28, 2017 from SnapGene 4.0.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5681
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     167..189
                     /label=fw to7000thiol
     primer_bind     468..483
                     /label=#177 FW DF1ORI
                     /note="#177 FW DF1ORI"
     primer_bind     503..538
                     /label=#213 REV AmpR
                     /note="#213 REV AmpR"
     CDS             complement(560..1375)
                     /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      1497..2085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2068..2085)
                     /label=#182 FW P15A
                     /note="#182 FW P15A"
     primer_bind     2088..2103
                     /label=#927 REV pA_Term
                     /note="#927 REV pA_Term"
     primer_bind     2091..2115
                     /label=#986 REV pA_Term #2
                     /note="#986 REV pA_Term #2"
     primer_bind     2147..2183
                     /label=#277 REV LIN#3
                     /note="#277 REV LIN#3"
     primer_bind     2147..2164
                     /label=#113 REV linear DNA
                     /note="#113 REV linear DNA"
     primer_bind     2152..2183
                     /label=#193 LINEAR 2
                     /note="#193 LINEAR 2"
     primer_bind     complement(2250..2266)
                     /label=#174 FW RSF
                     /note="#174 FW RSF"
     CDS             complement(2515..2706)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3515..4597)
                     /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"
     primer_bind     3551..3571
                     /label=#165 FW TRNA
                     /note="#165 FW TRNA"
     promoter        complement(4598..4675)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4866..4883
                     /label=#1123 T7FW#3
                     /note="#1123 T7FW#3"
     primer_bind     4960..4979
                     /label=#1 T7 Fw
                     /note="#1 T7 Fw"
     primer_bind     4960..4979
                     /label=T7 FW
     primer_bind     complement(4965..4983)
                     /label=#319 REV T7 Promoter
                     /note="#319 REV T7 Promoter"
     primer_bind     complement(4966..4988)
                     /label=#1082 rv from t7fw2
                     /note="#1082 rv from t7fw2"
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4986..5013)
                     /label=#1349 rv t7no2
                     /note="#1349 rv t7no2"
     primer_bind     4989..5011
                     /label=T7FW#2
     primer_bind     5003..5034
                     /label=#390 FW lac operator
                     /note="#390 FW lac operator"
     protein_bind    5003..5027
                     /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(5015..5074)
                     /label=#1057 REV pET28a w/o N Tags HOT
                     /note="#1057 REV pET28a w/o N Tags HOT"
     primer_bind     complement(5034..5080)
                     /label=#270 REV T7#2
                     /note="#270 REV T7#2"
     primer_bind     5039..5074
                     /label=#1617 FW_RBSMCS
                     /note="#1617 FW_RBSMCS"
     primer_bind     5040..5074
                     /label=#676 FW w/o N-tag pet28a
                     /note="#676 FW w/o N-tag pet28a"
     primer_bind     complement(5040..5074)
                     /label=#440 Rev w/o N-tag pET28a
                     /note="#440 Rev w/o N-tag pET28a"
     primer_bind     complement(5043..5073)
                     /label=#349 REV-w/o N-Tags MCS Met
                     /note="#349 REV-w/o N-Tags MCS Met"
     primer_bind     complement(5044..5080)
                     /label=rv RBS MGT
     primer_bind     complement(5044..5080)
                     /label=rv ATG to MAT
     primer_bind     complement(5045..5080)
                     /label=#250 REV T7 #2
                     /note="#250 REV T7 #2"
     primer_bind     complement(5045..5074)
                     /label=REV N-StrepII MCS
     primer_bind     complement(5045..5074)
                     /label=REV HIS
     primer_bind     complement(5048..5075)
                     /label=REV ybbR
     primer_bind     complement(5049..5080)
                     /label=#1108 rv noN-tags
                     /note="#1108 rv noN-tags"
     primer_bind     complement(5049..5080)
                     /label=rv noN-tags MGT
     primer_bind     5057..5080
                     /label=#130 FW Vio (MCS) GA
                     /note="#130 FW Vio (MCS) GA"
     primer_bind     complement(5057..5080)
                     /label=#112 REV T7
                     /note="#112 REV T7"
     primer_bind     complement(5057..5080)
                     /label=REV-w/o N-Tags MCS
     RBS             5058..5063
     primer_bind     complement(5063..5092)
                     /label=rv QC FgBetaF1
     CDS             5072..5533
                     /codon_start=1
                     /label=Feature 16
                     /translation="MGTNEEGFFSARGHRPLDGSGSGSGSAGTGSGADPEKSYAEGPGL
                     DGGESFQPSKFKIHAVDPDGVHRTDGGDGFVVTIEGPAPVDPVMVDNGDGTYDVEFEPK
                     EAGDYVINLTLDGDNVNGFPKTVTVKPAPSGHHHHHHGSDSLEFIASKLA"
     primer_bind     complement(5072..5095)
                     /label=rv QC FgB-F
     CDS             5072..5074
                     /codon_start=1
                     /label=ATG
                     /translation="M"
     CDS             5081..5125
                     /codon_start=1
                     /label=Fg beta
                     /translation="NEEGFFSARGHRPLD"
     primer_bind     5081..5104
                     /label=fw Fgb
     primer_bind     5096..5114
                     /label=fw QC FgB-F
     primer_bind     5099..5116
                     /label=#1485 fw QC FgbnoF
                     /note="#1485 fw QC FgbnoF"
     primer_bind     5099..5115
                     /label=fw Fgbshort
     primer_bind     5105..5148
                     /label=fw FgA(26-41) N
     primer_bind     5108..5125
                     /label=fw Fgb THR
     primer_bind     5125..5148
                     /label=fw QC GFFFGGG
     primer_bind     5125..5148
                     /label=fw 3FFFSARG
     primer_bind     5125..5148
                     /label=fw FFSARG
     primer_bind     5126..5149
                     /label=fw 4GS
     primer_bind     5126..5148
                     /label=fw Fgateb N
     primer_bind     5126..5147
                     /label=fw QC CFH
     primer_bind     complement(5149..5167)
                     /label=#1348 rv link fgbf3
                     /note="#1348 rv link fgbf3"
     CDS             5168..5470
                     /codon_start=1
                     /label=ddFLN4(C18S)
                     /translation="ADPEKSYAEGPGLDGGESFQPSKFKIHAVDPDGVHRTDGGDGFVV
                     TIEGPAPVDPVMVDNGDGTYDVEFEPKEAGDYVINLTLDGDNVNGFPKTVTVKPAP"
     primer_bind     5168..5191
                     /label=#1347 fw dd4_2
                     /note="#1347 fw dd4_2"
     primer_bind     complement(5450..5470)
                     /label=rv ddFLN4 linkdoc1
     primer_bind     complement(5450..5470)
                     /label=IV#567 rv ddFLN4 linker
                     /note="BoxIV#567 rv ddFLN4 linker"
     primer_bind     complement(5450..5470)
                     /label=IV#682 rv ddFLN4 8GSXdoc
                     /note="BoxIV#682 rv ddFLN4 8GSXdoc"
     primer_bind     complement(5450..5470)
                     /label=rv ddFLN4
     primer_bind     complement(5450..5470)
                     /label=rv ddFLN4 linker
     primer_bind     complement(5450..5470)
                     /label=rv ddFLN4 8GSXdoc
     primer_bind     5471..5557
                     /label=fw HIS-y-LPETGG
     primer_bind     5471..5496
                     /label=fw SG HIS
     CDS             5477..5494
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     5501..5556
                     /label=#107 FW T7 Term ybbr
                     /note="#107 FW T7 Term ybbr"
     CDS             5501..5533
                     /codon_start=1
                     /label=ybbR Tag
                     /translation="DSLEFIASKLA"
     primer_bind     complement(5501..5520)
                     /label=#871 REV ybbr (2)
                     /note="#871 REV ybbr (2)"
     primer_bind     5501..5520
                     /label=#75 FW ybbR
                     /note="#75 FW ybbR"
     primer_bind     5501..5520
                     /label=FW ybbR
     primer_bind     complement(5509..5533)
                     /label=#1616 rv fybbr Cterm
                     /note="#1616 rv fybbr Cterm"
     primer_bind     5520..5549
                     /label=FW QC Cys CTerm
     primer_bind     5534..5560
                     /label=#1575 fw_pET28
                     /note="#1575 fw_pET28"
     primer_bind     complement(5534..5557)
                     /label=#726 rv w/o CTags MCS#2
                     /note="#726 rv w/o CTags MCS#2"
     primer_bind     complement(5534..5557)
                     /label=rv w/o CTags MCS#2
     primer_bind     5534..5556
                     /label=FW-w/o C-Tags MCS
     primer_bind     complement(5534..5555)
                     /label=#1182 rv to STOP
                     /note="#1182 rv to STOP"
     primer_bind     complement(5534..5551)
                     /label=REV w/o C Tags
     primer_bind     5537..5556
                     /label=#109 FW T7 Term
                     /note="#109 FW T7 Term"
     primer_bind     complement(5587..5605)
                     /label=#769 rv to7000 (19-mer)
                     /note="#769 rv to7000 (19-mer)"
     primer_bind     complement(5587..5605)
                     /label=rv to7000
     primer_bind     complement(5591..5609)
                     /label=T7REV#2
     terminator      5609..5656
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5656..5679)
                     /label=#2 T7 Rev
                     /note="#2 T7 Rev"
     primer_bind     complement(5656..5679)
                     /label=T7 REV
     primer_bind     5676..11
                     /label=#897 FW dF1 ORI
                     /note="#897 FW dF1 ORI"
     primer_bind     complement(5676..11)
                     /label=#176 REV F1 ORI
                     /note="#176 REV F1 ORI"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct gtagaaataa
     5041 ttttgtttaa ctttaagaag gagatataca tatgggtacc aatgaagagg gctttttcag
     5101 cgcacgtggt catcgtccgc tggatggtag cggtagcggt tcaggtagcg caggtacagg
     5161 tagcggtgca gatccggaaa aaagctatgc cgaaggtccg ggtctggatg gtggtgaaag
     5221 ttttcagccg agcaaattca aaattcatgc agttgatccg gatggtgttc atcgtaccga
     5281 tggcggtgat ggttttgttg ttaccattga aggtcctgca ccggtggatc cggttatggt
     5341 tgataatggt gatggcacct atgatgttga atttgaaccg aaagaagcag gcgattacgt
     5401 tattaatctg accctggatg gcgataatgt taatggcttt ccgaaaaccg ttaccgttaa
     5461 accggcaccg agcggtcatc atcatcacca tcatggtagc gactctctgg aattcatcgc
     5521 ttctaaactg gcttaactcg agtaagatcc ggctgctaac aaagcccgaa aggaagctga
     5581 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     5641 cttgaggggt tttttgctga aaggaggaac tatatccgga t
//
