LOCUS Exported 9029 bp ds-DNA circular SYN 01-MAY-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pZac-mCherry-hPMCA2wb Upenn SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9029) AUTHORS Bal Khakh TITLE Direct Submission JOURNAL Exported Tuesday, May 1, 2018 from SnapGene Viewer 3.1.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9029 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 40..161 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" misc_feature 285..452 /note="ITR" primer_bind complement(471..487) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" rep_origin 629..1084 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 1110..1214 /gene="bla" /note="AmpR promoter" CDS 1215..2075 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 2246..2834 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 3122..3143 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." promoter 3158..3188 /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 3196..3212 /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)." primer_bind 3220..3236 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" misc_feature 3274..3441 /note="ITR" intron 4254..4386 /note="chimeric intron" /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" promoter 4431..4449 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 4455..5162 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /note="mCherry" /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" CDS 5178..5204 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /note="HA" /translation="YPYDVPDYA" CDS 5205..5222 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 5223..5240 /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" misc_feature 5298..9029 /note="PMCA2w/b" CDS complement(5799..5810) /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" ORIGIN 1 tctagagtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg 61 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta 121 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc 181 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct 241 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat 301 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg 361 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc 421 cgggcggcct cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc 481 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 541 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 601 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg 661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 721 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 781 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 841 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 901 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 961 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 1021 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca 1081 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 1141 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 1201 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 1261 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 1321 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 1381 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 1441 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 1501 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 1561 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 1621 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 1681 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 1741 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 1801 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 1861 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 1921 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 1981 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 2041 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 2101 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 2161 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 2221 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 2281 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 2341 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 2401 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 2461 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 2521 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 2581 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 2641 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 2701 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 2761 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 2821 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 2881 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 2941 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 3001 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 3061 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 3121 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 3181 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 3241 attacgccag atttaattaa ggccttaatt aggctgcgcg ctcgctcgct cactgaggcc 3301 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga 3361 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac 3421 ccgccatgct acttatctac gtagccatgc tctaggaaga tctaacatat cctggtgtgg 3481 agtaggggac gctgctctga cagaggctcg ggggcctgag ctggctctgt gagctgggga 3541 ggaggcagac agccaggcct tgtctgcaag cagacctggc agcattgggc tggccgcccc 3601 ccagggcctc ctcttcatgc ccagtgaatg actcaccttg gcacagacac aatgttcggg 3661 gtgggcacag tgcctgcttc ccgccgcacc ccagcccccc tcaaatgcct tccgagaagc 3721 ccattgagca gggggcttgc attgcacccc agcctgacag cctggcatct tgggataaaa 3781 gcagcacagc cccctagggg ctgcccttgc tgtgtggcgc caccggcggt ggagaacaag 3841 gctctattca gcctgtgccc aggaaagggg atcaggggat gcccaggcat ggacagtggg 3901 tggcaggggg ggagaggagg gctgtctgct tcccagaagt ccaaggacac aaatgggtga 3961 ggggagagct ctccccatag ctgggctgcg gcccaacccc accccctcag gctatgccag 4021 ggggtgttgc caggggcacc cgggcatcgc cagtctagcc cactccttca taaagccctc 4081 gcatcccagg agcgagcaga gccagagcag gttggagagg agacgcatca cctccgctgc 4141 tcgcaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg 4201 acacaacagt ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta 4261 tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa 4321 gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt gcctttctct 4381 ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact taatacgact 4441 cactataggc tagcatggtg agcaagggcg aggaggataa catggccatc atcaaggagt 4501 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg 4561 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg 4621 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg 4681 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct 4741 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact 4801 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct 4861 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt 4921 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg 4981 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg 5041 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg 5101 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca 5161 agtccggact cagatcctac ccctatgatg tgcctgacta tgcccaccac caccaccacc 5221 acctggtgcc caggggcagc agatctcgag ctcaagcttc gaattctgca gtcgacggta 5281 ccgcggtggc ggccgccatg ggtgacatga ccaacagcga cttttactcc aaaaaccaaa 5341 gaaatgagtc gagccatggg ggcgagttcg ggtgcacaat ggaggagctc cgctccctca 5401 tggagctgcg gggcactgag gctgtggtca agatcaagga gacttatggg gacaccgaag 5461 ccatctgccg gcgcctcaaa acctcacctg ttgaaggttt gccggacacc gctccagacc 5521 tggaaaagag aaagcaaatt tttgggcaaa actttatacc tccaaagaag ccaaaaacct 5581 tcctgcagct cgtgtgggag gcgctgcagg acgtgacgct catcatcctg gagattgccg 5641 ccatcatctc cctggggctg tccttctacc acccgcccgg cgagggcaac gaaggatgtg 5701 cgacggccca gggtggggca gaggatgaag gagaggcaga ggcaggttgg atcgaggggg 5761 ccgccattct cctctcagtt atctgtgtgg tcctggtcac ggccttcaat gactggagca 5821 aagagaaaca gttccggggc ctgcagagcc gcatcgagca ggaacagaaa tttaccgtag 5881 tccgggctgg ccaggtggtg cagatccctg tggctgagat cgtggttggg gacatagccc 5941 aggtcaaata tggtgacctc ctccctgccg acggcctctt catccagggc aatgacctca 6001 agattgatga aagctcccta actggagagt ctgaccaggt gcgcaagtcc gtggacaagg 6061 accccatgct gctgtcagga acccacgtga tggagggctc aggacggatg ttggtgactg 6121 ctgtgggtgt gaactctcag actggcatca tctttaccct cctgggggct ggtggtgaag 6181 aggaagagaa gaaagacaaa aaaggtgtga agaaggggga tggccttcag ctaccagcag 6241 cagacggtgc ggcagcttca aatgctgcag atagtgcgaa tgccagccta gtcaatggta 6301 aaatgcagga tggcaatgtg gacgccagcc agagcaaagc caaacagcag gacggggcag 6361 ccgccatgga gatgcagccc ctcaagagtg ccgagggcgg cgacgctgac gacaggaaga 6421 aggccagcat gcacaagaag gagaagtccg tgctgcaggg caagctcacc aagctggctg 6481 tgcagatcgg gaaggcgggc ttggtgatgt cagccatcac ggtgatcatc ctggtgctct 6541 acttcactgt ggacaccttc gtggtcaaca agaagccgtg gctgcctgag tgcacgcccg 6601 tctacgtgca gtactttgtc aagttcttca tcattggcgt gacggtgctg gtggtcgccg 6661 tgcccgaggg gctccctctg gccgtcacca tctcgttggc ctattcggtg aagaaaatga 6721 tgaaggacaa caacctggta cgccacctgg atgcctgtga gaccatgggc aatgccacag 6781 ccatctgctc agacaagaca ggcacgctga ccaccaatcg catgacagtg gtacaggcct 6841 atgtcggcga cgtccactat aaagagatcc ccgaccccag ctccatcaac accaagacca 6901 tggagctgct gatcaatgcc atcgccatca acagcgccta caccaccaag attctgcccc 6961 cagagaagga gggcgccctg cctcggcagg tgggcaacaa gacggagtgc ggcctgctgg 7021 gcttcgtgct ggacctgaag caggactacg agcccgtgcg cagccagatg ccagaggaga 7081 agttgtacaa agtgtacacc ttcaactccg tgcgcaagtc catgagcact gtcatcaagc 7141 tgcccgacga gagcttccgc atgtacagca agggggcttc tgagatcgtg ctcaagaagt 7201 gctgcaaaat cctcaatggg gcgggagagc ctcgtgtctt ccggccccgc gaccgggacg 7261 agatggtaaa gaaggtgatt gagcccatgg cttgcgatgg gctccgcact atctgcgtgg 7321 cctaccgcga cttccccagc agcccggagc cggactggga caatgagaat gacatcctca 7381 acgaactcac ctgcatctgc gtggtgggca tcgaggaccc ggtgcggcca gaggtcccag 7441 aagccatccg caagtgccag cgggcaggca tcacggtccg catggtcact ggcgacaata 7501 tcaacacggc tcgggccatc gccatcaagt gtggcatcat ccatcctggg gaggactttc 7561 tgtgcctcga gggcaaggag ttcaacagga ggatccgcaa cgagaagggg gagattgagc 7621 aggagcgaat tgacaagatc tggccaaagc tgcgggtgct ggctcgctcc tccccaacgg 7681 acaagcatac cctggttaaa ggcatcatcg acagcacaca cactgagcag cggcaggtgg 7741 tggccgtgac gggggacggg accaacgacg ggcctgcact caagaaggcc gacgtgggct 7801 tcgccatggg catcgcaggc actgacgtgg ccaaggaggc ctcagacatc atcctgacag 7861 acgacaattt cagcagcatc gtcaaggcag tgatgtgggg ccgcaacgtc tatgacagca 7921 tctccaaatt cttgcagttc cagctcaccg tcaacgtggt ggccgtgatt gtggccttca 7981 caggcgcctg catcacgcag gactcccctc tgaaggccgt gcagatgctc tgggtgaacc 8041 tcatcatgga cacgtttgcc tcgctggcac tggccactga gccgcccacg gagaccctgc 8101 tgctgaggaa gccgtacggc cgcaacaagc cgctcatctc caggaccatg atgaagaaca 8161 tcctgggcca tgctgtctac cagcttgccc tcatcttcac cctgctcttt gttggcgaga 8221 agatgttcca gatcgacagc gggaggaacg cgcccctgca ttcgccaccc tcagaacatt 8281 acaccatcat cttcaacacc ttcgtcatga tgcagctctt caacgagatc aacgcccgca 8341 agatccacgg cgagcgcaat gtctttgacg gcatcttccg gaaccccatc ttctgcacca 8401 tcgtgctggg cacctttgcc atccagatag tgatcgtgca gtttggaggg aagccattca 8461 gctgctctcc actgcagctg gaccagtgga tgtggtgcat attcattggg ttaggagagc 8521 tcgtttgggg ccaggtcatc gccaccatcc cgaccagcag actcaagttc ctcaaggagg 8581 caggcaggct cacacagaag gaggagatcc cggaggagga gctcaacgag gacgtggagg 8641 agatcgacca cgcggagcgg gagctgcggc ggggccagat cctgtggttc cgaggcctga 8701 atcggatcca gacacagatc cgcgtcgtga aggcgttccg tagctctctc tatgaaggtt 8761 tagaaaagcc tgaatctcga acctccatcc ataacttcat ggctcatcct gaattccgga 8821 tcgaagattc ccagccccac atccccctca ttgatgacac cgacctggaa gaagatgccg 8881 cgctcaagca gaactcgagc ccgccgtcat ccctcaacaa gaacaacagc gccatcgaca 8941 gtgggatcaa cctgacgacc gacacaagca aatcagctac ctcttcaagt ccagggagcc 9001 ccatccacag cctggagacg tcgctttag //