LOCUS       Exported                4919 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  RPU standard plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pAN1717
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4919)
  AUTHORS   Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski
            EA, Ross D, Densmore D, Voigt CA
  TITLE     Genetic circuit design automation.
  JOURNAL   Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341.
  PUBMED    27034378
REFERENCE   2  (bases 1 to 4919)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4919
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        114..163
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             213..932
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(258..279)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(519..538)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     866..887
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1080..2162
                     /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     complement(1099..1118)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2181..2804
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2254..2273)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2987..3006)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(3371..3915)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3451..3470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4004..4819)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4727..4746
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4811..4830
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttgat
       61 aagtccctaa cttttacagc tagctcagtc ctaggtatta tgctagcctg aagctgtcac
      121 cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac aaataatttt
      181 gtttaatact agagaaagag gggaaatact agatggtgag caagggcgag gagctgttca
      241 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      301 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      361 ccacaggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc
      421 aatgcttcgc ccgctacccc gaccacatga agctgcacga cttcttcaag tccgccatgc
      481 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      541 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      601 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      661 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
      721 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac accccaatcg
      781 gcgacggccc cgtgctgctg cccgacaacc actaccttag ctaccagtcc gccctgagca
      841 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      901 tcactctcgg catggacgag ctgtacaagt aactcggtac caaattccag aaaagaggcc
      961 tcccgaaagg ggggcctttt ttcgttttgg tccaatggcg gcgcgccatc gaatggcgca
     1021 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaata
     1081 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     1141 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     1201 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     1261 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     1321 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     1381 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     1441 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     1501 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     1561 tctcccatga ggacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     1621 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     1681 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     1741 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     1801 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     1861 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gatagctcat
     1921 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     1981 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgccag
     2041 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     2101 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2161 gataatccag gaggaaaaaa atgtccagat tagataaaag taaagtgatt aacagcgcat
     2221 tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc
     2281 taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg
     2341 ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa
     2401 gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc
     2461 gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg
     2521 aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac
     2581 tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg
     2641 ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta
     2701 tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca
     2761 tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ctaataattg gtaacgaatc
     2821 agacaattga cggctcgagg gagtagcata gggtttgcag aatccctgct tcgtccattt
     2881 gacaggcaca ttatgcatcg atgataagct gtcaaacatg agcagatcct ctacgccgga
     2941 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     3001 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcaaata ttttatctga
     3061 ggtgcttcct cgctcactga ctcgctgcac gaggcagacc tcagcgctag cggagtgtat
     3121 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     3181 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     3241 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     3301 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     3361 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     3421 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc ctggcggctc
     3481 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     3541 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     3601 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     3661 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     3721 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     3781 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     3841 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     3901 ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg acgctcagtg gaacgaaaaa
     3961 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accttagaaa aactcatcga
     4021 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     4081 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     4141 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     4201 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     4261 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     4321 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     4381 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     4441 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     4501 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     4561 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     4621 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     4681 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     4741 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag caagacgttt
     4801 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     4861 ttgttcatga tgatatattt ttatcttgtg caatgtacat cagagatttt gagacacaa
//
