LOCUS Exported 7272 bp ds-DNA circular SYN 23-SEP-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3 Flag beta-2-adrenergic-receptor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7272) AUTHORS Tang Y, Hu LA, Miller WE, Ringstad N, Hall RA, Pitcher JA, DeCamilli P, Lefkowitz RJ TITLE Identification of the endophilins (SH3p4/p8/p13) as novel binding partners for the beta1-adrenergic receptor. JOURNAL Proc Natl Acad Sci U S A. 1999 Oct 26. 96(22):12559-64. PUBMED 10535961 REFERENCE 2 (bases 1 to 7272) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7272 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(63..80) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(314..333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(993..1853) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1616..1635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1854..1958) /gene="bla" /label=AmpR promoter primer_bind complement(2033..2052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2224..2603 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2604..2807 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2758..2778 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2852..2871 /label=T7 /note="T7 promoter, forward primer" promoter 2852..2870 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(4814..4832) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4815..4832) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4852..4869) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4858..5082 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5128..5556 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5215..5234) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5425..5446 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5565..5585) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5570..5899 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5750..5885 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5812..5831 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5966..6760 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6020..6039) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6630..6649 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6934..7055 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6971..6990) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7025..7044 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7104..7120) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7104..7120) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7117..7139) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7128..7144 /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)." promoter complement(7152..7182) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7197..7218 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 721 aaaatagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 2881 cttggtaccg agctcggatc caccatgaag accatcatcg ccctgagcta catcttctgc 2941 ctggtgttcg ccgactacaa ggacgatgat gacgccgggc aacccgggaa cggcagcgcc 3001 ttcttgctgg cacccaatag aagccatgcg ccggaccacg acgtcacgca gcagagggac 3061 gaggtgtggg tggtgggcat gggcatcgtc atgtctctca tcgtcctggc catcgtgttt 3121 ggcaatgtgc tggtcatcac agccattgcc aagttcgagc gtctgcagac ggtcaccaac 3181 tacttcatca cttcactggc ctgtgctgat ctggtcatgg gcctggcagt ggtgcccttt 3241 ggggccgccc atattcttat gaaaatgtgg acttttggca acttctggtg cgagttttgg 3301 acttccattg atgtgctgtg cgtcacggcc agcattgaga ccctgtgcgt gatcgcagtg 3361 gatcgctact ttgccattac ttcacctttc aagtaccaga gcctgctgac caagaataag 3421 gcccgggtga tcattctgat ggtgtggatt gtgtcaggcc ttacctcctt cttgcccatt 3481 cagatgcact ggtaccgggc cacccaccag gaagccatca actgctatgc caatgagacc 3541 tgctgtgact tcttcacgaa ccaagcctat gccattgcct cttccatcgt gtccttctac 3601 gttcccctgg tgatcatggt cttcgtctac tccagggtct ttcaggaggc caaaaggcag 3661 ctccagaaga ttgacaaatc tgagggccgc ttccatgtcc agaaccttag ccaggtggag 3721 caggatgggc ggacggggca tggactccgc agatcttcca agttctgctt gaaggagcac 3781 aaagccctca agacgttagg catcatcatg ggcactttca ccctctgctg gctgcccttc 3841 ttcatcgtta acattgtgca tgtgatccag gataacctca tccgtaagga agtttacatc 3901 ctcctaaatt ggataggcta tgtcaattct ggtttcaatc cccttatcta ctgccggagc 3961 ccagatttca ggattgcctt ccaggagctt ctgtgcctgc gcaggtcttc tttgaaggcc 4021 tatgggaatg gctactccag caacggcaac acaggggagc agagtggata tcacgtggaa 4081 caggagaaag aaaataaact gctgtgtgaa gacctcccag gcacggaaga ctttgtgggc 4141 catcaaggta ctgtgcctag cgataacatt gattcacaag ggaggaattg tagtacaaat 4201 gactcactgc tgtaaagcag tttttctact tttaaagacc ccccccccca acagaacact 4261 aaacagacta tttaacttga gggtaataaa cttagaataa aattgtaaaa ttgtatagag 4321 atatgcagaa ggaagggcat ccttctgcct tttttatttt tttaagctgt aaaaagagag 4381 aaaacttatt tgagtgatta tttgttattt gtacagttca gttcctcttt gcatggaatt 4441 tgtaagttta tgtctaaaga gctttagtcc tagaggacct gagtctgcta tattttcatg 4501 acttttccat gtatctgcct cactattcaa gtattagggg taatatattg ctgctggtaa 4561 tttgtatctg aaggagattt tccttcctac acccttggac ttgaggattt tgagtatctc 4621 ggacctttca gctgtgaaca tggactcttc ccccactcct cttatttgct cacacggggt 4681 attttaggca gggatttgag gagcagcttc agttgttttc ccgagcaaag tctaaagttt 4741 acagtaaata aattgtttga ccatgaaaaa aaaaaaaaag gtagcagtcg agcatgcatc 4801 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg 4861 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 4921 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 4981 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 5041 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa 5101 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg 5161 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 5221 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 5281 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 5341 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 5401 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 5461 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 5521 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt 5581 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 5641 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 5701 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 5761 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 5821 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 5881 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca 5941 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 6001 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 6061 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 6121 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 6181 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 6241 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 6301 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 6361 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 6421 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 6481 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 6541 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 6601 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 6661 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 6721 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 6781 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 6841 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 6901 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 6961 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 7021 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 7081 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 7141 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 7201 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 7261 tcgtgccagc tg //