LOCUS Exported 7413 bp ds-DNA circular SYN 20-FEB-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pHJ12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7413) AUTHORS Trial User TITLE Direct Submission JOURNAL Exported 20 Feb 2018 from SnapGene 2.6.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7413 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(294..1010) /codon_start=1 /note="sfGFP" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" RBS complement(1011..1022) /note="rbs K143021" promoter complement(1029..1190) /note="PcadBA" terminator 1238..1265 /note="T7Te terminator" /note="phage T7 early transcription terminator" promoter 1366..1420 /note="pLacO1" protein_bind 1368..1384 /bound_moiety="lac repressor encoded by lacI" /note="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)." protein_bind 1391..1407 /bound_moiety="lac repressor encoded by lacI" /note="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)." RBS 1427..1450 /note="RBS benchmarking" CDS 1451..2425 /codon_start=1 /note="CadC-VHH-Caffeine (No linker)" /translation="MQQPVVRVGEWLVTPSINQISRNGRQLTLEPRLIDLLVFFAQHSG EVLSRDELIDNVWKRSIVTNHVVTQSISELRKSLKDNDEDSPVYIATVPKRGYKLMVPV IWYSEEEGEEIMLSSPPPIPEAVPATDSPSHSLNIQNTATPPEQSPVKSKRGGPGLLLL LLLLLLLLLLLLGPGGEVQLQASGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQAPGK EREFLATVGWSSGITYYMDSVKGRFTISRDNAKNSAYLQMNSLKPEDTAVYYCTATRAY SVGYDYWGQGTQVTVSHAAAGAPVPYPDPLEPREQKLISEEDL" CDS 2393..2422 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /note="Myc" /translation="EQKLISEEDL" terminator 2520..2591 /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2607..2634 /note="T7Te terminator" /note="phage T7 early transcription terminator" operon 2681..3957 /operon="lacI" /note="lacI" /note="lacI operon" CDS 2875..3957 /codon_start=1 /gene="lacI" /product="lac repressor" /note="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 EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" terminator 4073..4104 /note="tonB terminator" /note="bidirectional E. coli tonB-P14 transcription terminator" CDS complement(4203..5018) /codon_start=1 /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" CDS complement(5376..6326) /codon_start=1 /gene="rep101" /product="protein needed for replication with the pSC101 origin" /note="Rep101" /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 6374..6596 /note="pSC101 ori" /note="low-copy replication origin that requires the Rep101 protein" terminator 7088..7182 /note="lambda t0 terminator" /note="transcription terminator from phage lambda" terminator 7338..7367 /note="T3Te terminator" /note="phage T3 early transcription terminator" ORIGIN 1 ctcggatacc cttactctgt tgaaaacgaa tagataggtt tgctcgtagt ttaccacgga 61 tacagacagt gataatctta ggcgcgccga atcttgcttg gcaaggttct tttttttttt 121 caaaaaagca cgaaaaaagc acccataagg taaacttttg agtgctttta aatacgcttc 181 acagtttctt cttcattttt aacaaaaaac ccagctcatt gagctgggtt taagctggct 241 tggcggcgtc ctaagattac tactgataac cactgttgat tgggataccc gtattattta 301 taaagttcgt ccataccgtg agtgatacct gctgctgtaa caaattcaag aagtaccata 361 tggtcacgtt tttcgttagg atcttttgaa agaactgatt gagtagaaag gtagtggtta 421 tctggaagca atactggtcc gtctccgatt ggtgtatttt gttgataatg atcagccaat 481 tgaacagatc catcttcaac attgtgacga attttaaaat tagctttaat accgtttttt 541 tgtttgtcag ctgtgatata aacattatga gagttgaagt tgtattcaag tttgtgtccc 601 aaaatgttac catcttcttt gaaatcgatt cctttcaatt cgatacgatt aacaagagtg 661 tcaccttcga attttacttc agcacgtgtt ttatatgttc cgtcatcttt aaatgaaatt 721 gtacgttctt gtacataacc ttctggcata gcagatttaa aaaaatcgtg acgtttcata 781 tgatcaggat aacgtgaaaa acattgtact ccatatgtca atgttgtaac aagagtaggc 841 caaggaacag gaagttttcc agttgtacaa atgaatttaa gtgtcaattt accgttagtt 901 gcatcacctt caccttcacc acgtacagaa aatttgtgac cgttaacatc accatccaat 961 tcaaccaaga taggtactac acctgtaaaa agttcttctc cttttgacat ttcaccacct 1021 ttactagtcc aaactggcga taagattact cacgaaaaaa ggattaatcc taaagattag 1081 gtgaaataac acaaaagttt ctgtaagtga gaacttgagg ttttttatta acacatcagg 1141 atcgcaagtt gatatcatga aaagataaac atttaatgtt tacaatggat aaggaacggt 1201 tatttctgcg tagatctatc ttacacagca tcacactggc tcaccttcgg gtgggccttt 1261 ctgcgtttat atactagaga gagaatataa aaagccagat tattaatccg gcttttttat 1321 tatttaggca actgaaacga ttcggatcct gtattactat tcttaaattg tgagcggata 1381 acaattgaca ttgtgagcgg ataacaagat actgagcaca gctagcgatt aactaataag 1441 gaggacaaac atgcaacaac ctgtagttcg cgttggcgaa tggcttgtta ctccgtccat 1501 aaaccaaatt agccgcaatg ggcgtcaact tacccttgag ccgagattaa tcgatcttct 1561 ggttttcttt gctcaacaca gtggcgaagt acttagcagg gatgaactta tcgataatgt 1621 ctggaagaga agtattgtca ccaatcacgt tgtgacgcag agtatctcag aactacgtaa 1681 gtcattaaaa gataatgatg aagatagtcc tgtctatatc gctactgtac caaagcgcgg 1741 ctataaatta atggtgccgg ttatctggta cagcgaagaa gagggagagg aaataatgct 1801 atcttcgcct ccccctatac cagaggcggt tcctgccaca gattctccct cccacagtct 1861 taacattcaa aacaccgcaa cgccacctga acaatcccca gttaaaagca aacgaggtgg 1921 cccaggtctg ttgctgttgc tgttgctgtt gctgttgctg ttgctgttgc tgttgggtcc 1981 aggtggcgaa gttcaactgc aagcttctgg cggcggcctg gttcaagctg gcggctctct 2041 gcgtctgtct tgcactgctt ctggccgtac tggcactatc tactctatgg cttggttccg 2101 tcaagctcct ggcaaagaac gtgaattcct ggctactgtt ggctggtctt ctggcatcac 2161 ttactacatg gattctgtta aaggccgttt cactatctct cgtgataacg ctaaaaactc 2221 tgcttacctg caaatgaact ctctgaaacc tgaagatact gctgtttact actgcactgc 2281 tactcgtgct tactctgttg gctacgatta ctggggccaa ggcactcaag ttactgtttc 2341 tcatgctgct gctggcgctc ctgttcctta ccctgatcct ctggaacctc gtgaacaaaa 2401 actgatctct gaagaagatc tgtaaacttt atctgagaat agtcaatctt cggaaatccc 2461 aggtggcatg ctaaaagtct cgtaaagcgt tctatcaata acccgttggt gccaggcatc 2521 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg 2581 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata 2641 ccgtctcaga atcggccgtg aacaataaaa tagtttcggt ttgcatttaa atcttacata 2701 tgtaatactt tcaaagacta catttgtaag atttgatgtt tgagtcggct gaaagatcgt 2761 acgtaccaat tattgtttcg tgattgttca agccataaca ctgtagggat agtggaaaga 2821 gtgcttcatc tggttacgat caatcaaata ttcaaacgga gggagacgat tttgatgaaa 2881 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc 2941 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg 3001 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg 3061 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg 3121 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc 3181 ggcgtcgaag cctgtaaaac ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg 3241 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat 3301 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc 3361 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc 3421 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat 3481 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc 3541 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg 3601 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg 3661 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat 3721 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac 3781 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca 3841 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 3901 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgataa 3961 taaaaggtcc cgtctgaact tactgtgaat tcgactaatt attgaccact tccgagtaga 4021 atcgtgcttc agtaagagag tcactaaggg ttagttagtt agattagcag aaagtcaaaa 4081 gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg ggctcactca 4141 aaggcggtaa tcagataaaa aaaatcctta gctttcgcta aggatgattt ctgctagtat 4201 tattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca 4261 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc 4321 caaagaatgg caaggtcctg gtaacggtct gcgattccga cccgtccaac atcaatacaa 4381 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg 4441 actgaatccg gtgagaatgg caagagcttg tgcatttctt tccagacttg ttcaacaggc 4501 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt catgcgtgat 4561 tgcgcctgag caagacgaaa tacacgatcg ctgttaaaag gacaattaca aacaggaatc 4621 gaatgtaacc ggcgcaggaa cacggccagc gcatcaacaa tattttcacc tgaatcagga 4681 tattcttcta atacctggaa ggctgttttc ccaggaatcg cggtggtgag taaccacgca 4741 tcatcaggag tacggataaa atgcttgatg gtcgggagag gcataaactc cgtcagccag 4801 ttgagacgga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga 4861 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg 4921 acattatcgc gagcccattt atacccatat aaatcagcgt ccatgttgga gtttaagcgc 4981 ggacgggagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt 5041 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat 5101 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag 5161 gatcagctct agtagttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa 5221 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg 5281 acagttttcc ctttgatatc taacggtgaa cagttgttct acttttgttt gttagtcttg 5341 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt 5401 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca 5461 tgcttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca 5521 gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg 5581 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga 5641 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta 5701 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc 5761 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat 5821 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat 5881 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta 5941 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat 6001 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta 6061 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat 6121 acaccataag cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac 6181 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag 6241 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt 6301 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta 6361 taccaattga gatgggctag tcaatgataa ttacatgtcc ttttcctttg agttgtgggt 6421 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta 6481 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga 6541 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta 6601 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa 6661 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg 6721 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca 6781 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt 6841 atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg 6901 gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct 6961 gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg 7021 ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag 7081 tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca 7141 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgtaaact 7201 tggtctgaca gctctagctc cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt 7261 tctttaaaac cgaaaagatt acttcgcgtt tgccacctga cgtctaagaa aaggaatatt 7321 cagcaatttg cccgtgccga agaaaggccc acccgtgaag gtgagccagt gagttgattg 7381 ctacgtaatt agttagttag cccttagtga ctc //