LOCUS Exported 5706 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTrcHisA-rat-PTP1B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5706) AUTHORS Cortesio CL, Chan KT, Perrin BJ, Burton NO, Zhang S, Zhang ZY, Huttenlocher A TITLE Calpain 2 and PTP1B function in a novel pathway with Src to regulate invadopodia dynamics and breast cancer cell invasion. JOURNAL J Cell Biol. 2008 Mar 10;180(5):957-71. PUBMED 18332219 REFERENCE 2 (bases 1 to 5706) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5706 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 193..222 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 230..246 /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 235..257 /label=M13/pUC Reverse /note="In lacZ gene" misc_feature 263..332 /label=rrnG antiterminator /note="antiterminator from the E. coli rrnG leader region (Berg et al., 1989)" CDS 383..409 /codon_start=1 /product="synthetic cistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream cistron (Schoner, 1997)" /label=minicistron /note="This first cistron should terminate 3 bp upstream of the ATG for the second cistron." /translation="MYRLNKEE" CDS 425..442 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 446..478 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 491..505 /codon_start=1 /product="enterokinase recognition and cleavage site" /label=enterokinase site /translation="DDDDK" primer_bind complement(1889..1906) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2059..2145 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2237..2264 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2284..2375 /gene="bla" /label=AmpR promoter CDS 2376..3236 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2594..2613) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3407..3995 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3896..3915 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4149..4166 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4181..4321 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4247..4266) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4407..4429 /label=pGEX 3' /note="pGEX vectors, reverse primer" promoter 4507..4584 /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 4585..5667 /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 YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(4604..4623) /label=LacI-R /note="LacI, reverse primer" ORIGIN 1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc 61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc 121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc 181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga 241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa 301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta 361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg 421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg 481 gactctgtac gacgatgacg ataaggatcg atggggatcc atggaaatgg agaaggaatt 541 cgagcagatc gataaggctg ggaactgggc ggctatttac caggatattc gacatgaagc 601 cagtgacttc ccatgcagaa tagcgaaact tcctaagaac aaaaaccgga acaggtaccg 661 agatgtcagc ccttttgacc acagtcggat taaattgcat caggaagata atgactatat 721 caatgccagc ttgataaaaa tggaggaagc ccagaggagc tatatcctca cccagggccc 781 tttaccaaac acgtgcgggc acttctggga gatggtgtgg gagcagaaga gcaggggcgt 841 ggtcatgctc aaccgcatca tggagaaagg ctcgttaaaa tgtgcccagt attggccaca 901 gaaagaagaa aaagagatgg tcttcgatga caccaatttg aagctgacac tgatctctga 961 agatgtcaag tcatattaca cagtacggca gttggagttg gagaacctgg ctacccagga 1021 ggctcgagag atcctgcatt tccactacac cacctggcct gactttggag tccctgagtc 1081 acctgcctct ttcctcaatt tcctattcaa agtccgagag tcaggctcac tcagcccaga 1141 gcacggcccc attgtggtcc actgcagtgc tggcattggc aggtcaggga ccttctgcct 1201 ggctgacacc tgcctcttac tgatggacaa gaggaaagac ccgtcctctg tggacatcaa 1261 gaaagtgctg ttggagatgc gcaggttccg catggggctc atccagacgg ccgaccaact 1321 gcgcttctcc tacctggctg tgatcgaggg tgcaaagttc atcatgggcg actcgtcagt 1381 gcaggatcag tggaaggagc tttcccatga agacctggag cctccccctg agcacgtgcc 1441 cccacctccc cggccaccca aacgcacatt ggagcctcac aatggcaagt gcaaggagct 1501 cttctccaac caccagtggg tgagcgagga gagctgtgag gatgaggaca tcctggccag 1561 agaggaaagc agagccccct caattgctgt gcacagcatg agcagtatga gtcaagacac 1621 tgaagttagg aaacggatgg tgggtggagg tcttcaaagt gctcaggcat ctgtccccac 1681 tgaggaagag ctgtccccaa ccgaggagga acaaaaggca cacaggccag ttcactggaa 1741 gcccttcctg gtcaacgtgt gcatggccac ggccctggcg actggcgcgt acctctgtta 1801 ccgggtatgt tttcactgaa gatctgcagc tggtaccata tgggaattcg aagcttggct 1861 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg 1921 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc 1981 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag 2041 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt 2101 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat 2161 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc 2221 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact 2281 ctttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 2341 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 2401 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 2461 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 2521 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 2581 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac 2641 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 2701 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 2761 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 2821 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 2881 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc 2941 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 3001 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 3061 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc 3121 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 3181 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 3241 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 3301 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 3361 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 3421 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 3481 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 3541 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 3601 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 3661 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 3721 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 3781 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 3841 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 3901 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 3961 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 4021 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 4081 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 4141 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc 4201 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg 4261 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc 4321 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc 4381 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc 4441 atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt 4501 tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa 4561 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat 4621 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg 4681 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg 4741 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc 4801 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc 4861 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac 4921 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat 4981 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac 5041 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag 5101 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc 5161 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg 5221 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg 5281 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc 5341 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga 5401 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat 5461 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg 5521 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc 5581 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 5641 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt 5701 gatctg //