LOCUS       62056_7800        5925 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5925)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..81
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     protein_bind    84..100
                     /label=lac operator
                     /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)."
     5'UTR           122..255
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     CDS             256..270
                     /codon_start=1
                     /label=TEE
                     /note="translation enhancing element for E. coli (Qing et
                     al., 2004)"
                     /translation="MNHKV"
     CDS             271..288
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             289..300
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             922..1869
                     /codon_start=1
                     /label=iRFP720
                     /note="near-infrared fluorescent protein with an emission
                     peak at 720 nm, engineered from a bacterial phytochrome 
                     (Shcherbakova and Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     3'UTR           1886..2030
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     rep_origin      complement(2229..2684)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2784..2888
                     /label=AmpR promoter
     CDS             2889..3746
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3920..4508
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(4646..4667)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4683..5762)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5763..5840)
                     /label=lacI promoter
ORIGIN
        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
      241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcatat cgaaggtagg
      301 catatggagc tcggtacctc tagcggtgct cagtacgaag atggtaaaca gtacactacc
      361 ctggaaaaac cggtagctgg cgcgccgcaa gtgctggagt ttttctcttt cttcagcccg
      421 cattcctatc agtttgaaga agttctgcat atttctgata atgtgaagaa aaaactgccg
      481 gaaggcgtga agatgactaa ataccacgtc aacttcatgg gtggtgacct gggcaaagat
      541 ctgactcagg catgggctgt ggcgatggcg ctgggcgtgg aagacaaagt gactgttccg
      601 ctgtttgaag gcgtacagaa aacccagacc attcgttctg cttctgatat ccgcgatgta
      661 tttatcaacg caggtattaa aggtgaagag tacgacgcgg cgtggaacag cttcgtggtg
      721 aaatctctgg tcgctcagca ggaaaaagct gcagctgacg tgcaattgcg tggcgttccg
      781 gcgatgtttg ttaacggtaa atatcagctg aatccgcagg gtatggatac cagcaatatg
      841 gatgtttttg ttcagcagta tgctgataca gtgaaatatc tgtccgagaa aaaaggtgat
      901 gaagatgaag atagcggatc catggcggaa ggatccgtcg ccaggcagcc tgacctcttg
      961 acctgcgacg atgagccgat ccatatcccc ggtgccatcc aaccgcatgg actgctgctc
     1021 gccctcgccg ccgacatgac gatcgttgcc ggcagcgaca accttcccga actcaccgga
     1081 ctggcgatcg gcgccctgat cggccgctct gcggccgatg tcttcgactc ggagacgcac
     1141 aaccgtctga cgatcgcctt ggccgagccc ggggcggccg tcggagcacc gatcactgtc
     1201 ggcttcacga tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca tgatcagctc
     1261 atcttcctcg agctcgagcc tccccagcgg gacgtcgccg agccgcaggc gttcttccgc
     1321 cgcaccaaca gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc
     1381 gccgcggcgc aagaggtgcg gaagattacc ggcttcgatc gggtgatgat ctatcgcttc
     1441 gcctccgact tcagcgggtc cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa
     1501 ctaggcctgc actatcctgc ctcattcatc ccggcgcagg cccgtcggct ctataccatc
     1561 aacccggtac ggatcattcc cgatatcaat tatcggccgg tgccggtcac cccagacctc
     1621 aatccggtca ccgggcggcc gattgatctt agcttcgcca tcctgcgcag cgtctcgccc
     1681 aaccatctgg agttcatgcg caacataggc atgcacggca cgatgtcgat ctcgattttg
     1741 cgcggcgagc gactgtgggg attgatcgtt tgccatcacc gaacgccgta ctacgtcgat
     1801 ctcgatggcc gccaagcctg cgagctagtc gcccaggttc tggcctggca gatcggcgtg
     1861 atggaagagt aatctagata ggtaatctct gcttaaaagc acagaatcta agatccctgc
     1921 catttggcgg ggattttttt atttgttttc aggaaataaa taatcgatcg cgtaataaaa
     1981 tctattatta tttttgtgaa gaataaattt gggtgcaatg agaatgcgca ggccctttcg
     2041 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2101 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2161 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     2221 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2281 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2341 gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2401 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2461 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2521 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2581 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2641 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     2701 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgtcag
     2761 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2821 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2881 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2941 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3001 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3061 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3121 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3181 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3241 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3301 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3361 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3421 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3481 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3541 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3601 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3661 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3721 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3781 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3841 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4561 ctcacatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4621 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4741 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     4801 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     4861 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     4921 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     4981 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5041 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5101 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5161 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5221 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5281 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5341 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5401 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5461 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5521 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5581 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5641 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5701 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     5761 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     5821 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     5881 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgc
//
