LOCUS Exported 9435 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS BC-GPR56 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9435) TITLE BC-GPR56 JOURNAL Unpublished REFERENCE 2 (bases 1 to 9435) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9435 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 580..921 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 988..1404 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1334..1356 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1372..1388 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 3214..3223 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 3656..3673 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3657..3673 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3680..3699 /label=T7 /note="T7 promoter, forward primer" promoter 3680..3698 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 3749..4210 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3805..3822) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4032..4051 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4251..4847 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4251..4270) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4747..4767 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 5748..6263 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" primer_bind complement(6432..6448) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6432..6448) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6445..6467) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6456..6472 /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(6480..6510) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6525..6546 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6663..6680) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6834..7422) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6914..6933) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7593..8453) /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 8216..8235 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8454..8558) /gene="bla" /label=AmpR promoter primer_bind 8626..8644 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8682..8704) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8804..8823 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9215..9234 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 9434..516 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt 301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc 361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat 421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc 481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag 541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc 601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg 661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga 721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg 781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga 841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga 901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact 961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc 1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag 1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga 1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc 1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac 1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc 1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca 1381 gccctcactc cttctctagg cgccggaaat tagatctacc ggtaagagtg actccgtcgg 1441 aggaaaatga ctccccagtc gctgctgcag acgacactgt tcctgctgag tctgctcttc 1501 ctggtccaag gtgcccacgg caggggccac agggaagact ttcgcttctg cagccagcgg 1561 aaccagacac acaggagcag cctccactac aaacccacac cagacctgcg catctccatc 1621 gagaactccg aagaggccct cacagtccat gcccctttcc ctgcagccca ccctgcttcc 1681 cgatccttcc ctgaccccag gggcctctac cacttctgcc tctactggaa ccgacatgct 1741 gggagattac atcttctcta tggcaagcgt gacttcttgc tgagtgacaa agcctctagc 1801 ctcctctgct tccagcacca ggaggagagc ctggctcagg gccccccgct gttagccact 1861 tctgtcacct cctggtggag ccctcagaac atcagcctgc ccagtgccgc cagcttcacc 1921 ttctccttcc acagtcctcc ccacacggcc gctcacaatg cctcggtgga catgtgcgag 1981 ctcaaaaggg acctccagct gctcagccag ttcctgaagc atccccagaa ggcctcaagg 2041 aggccctcgg ctgcccccgc cagccagcag ttgcagagcc tggagtcgaa actgacctct 2101 gtgagattca tgggggacat ggtgtccttc gaggaggacc ggatcaacgc cacggtgtgg 2161 aagctccagc ccacagccgg cctccaggac ctgcacatcc actcccggca ggaggaggag 2221 cagagcgaga tcatggagta ctcggtgctg ctgcctcgaa cactcttcca gaggacgaaa 2281 ggccggagcg gggaggctga gaagagactc ctcctggtgg acttcagcag ccaagccctg 2341 ttccaggaca agaattccag ccacgtcctg ggtgagaagg tcttggggat tgtggtacag 2401 aacaccaaag tagccaacct cacggagccc gtggtgctca ccttccagca ccagctacag 2461 ccgaagaatg tgactctgca atgtgtgttc tgggttgaag accccacatt gagcagcccg 2521 gggcattgga gcagtgctgg gtgtgagacc gtcaggagag aaacccaaac atcctgcttc 2581 tgcaaccact tgacctactt tgcagtgctg atggtctcct cggtggaggt ggacgccgtg 2641 cacaagcact acctgagcct cctctcctac gtgggctgtg tcgtctctgc cctggcctgc 2701 cttgtcacca ttgccgccta cctctgctcc agggtgcccc tgccgtgcag gaggaaacct 2761 cgggactaca ccatcaaggt gcacatgaac ctgctgctgg ccgtcttcct gctggacacg 2821 agcttcctgc tcagcgagcc ggtggccctg acaggctctg aggctggctg ccgagccagt 2881 gccatcttcc tgcacttctc cctgctcacc tgcctttcct ggatgggcct cgaggggtac 2941 aacctctacc gactcgtggt ggaggtcttt ggcacctatg tccctggcta cctactcaag 3001 ctgagcgcca tgggctgggg cttccccatc tttctggtga cgctggtggc cctggtggat 3061 gtggacaact atggccccat catcttggct gtgcatagga ctccagaggg cgtcatctac 3121 ccttccatgt gctggatccg ggactccctg gtcagctaca tcaccaacct gggcctcttc 3181 agcctggtgt ttctgttcaa catggccatg ctagccacca tggtggtgca gatcctgcgg 3241 ctgcgccccc acacccaaaa gtggtcacat gtgctgacac tgctgggcct cagcctggtc 3301 cttggcctgc cctgggcctt gatcttcttc tcctttgctt ctggcacctt ccagcttgtc 3361 gtcctctacc ttttcagcat catcacctcc ttccaaggct tcctcatctt catctggtac 3421 tggtccatgc ggctgcaggc ccggggtggc ccctcccctc tgaagagcaa ctcagacagc 3481 gccaggctcc ccatcagctc gggcagcacc tcgtccagcc gcatctagaa gggcgaattc 3541 tgcagatatc catcacactg gcggccgcag cacagtggtc gaaattctgg cgaaaggggg 3601 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa 3661 aacgacggcc agtgaattgt aatacgactc actatagggc gaattaattc cggttatttt 3721 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga 3781 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg 3841 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt 3901 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat 3961 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg 4021 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg 4081 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt 4141 cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac 4201 acgatgataa tatggccacc acccatatga aagaaaccgc tgctgctaaa atgaccgagt 4261 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg 4321 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg 4381 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 4441 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 4501 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 4561 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 4621 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 4681 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 4741 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 4801 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctgg tcgacaagca 4861 tggggtaaag tactgttctc atcacatcat atcaaggtta tataccatca atattgccac 4921 agatgttact tagcctttta atatttctct aatttagtgt atatgcaatg atagttctct 4981 gatttctgag attgagtttc tcatgtgtaa tgattattta gagtttctct ttcatctgtt 5041 caaatttttg tctagtttta ttttttactg atttgtaaga cttcttttta taatctgcat 5101 attacaattc tctttactgg ggtgttgcaa atattttctg tcattctatg gcctgacttt 5161 tcttaatggt tttttaattt taaaaataag tcttaatatt catgcaatct aattaacaat 5221 cttttctttg tggttaggac tttgagtcat aagaaatttt tctctacact gaagtcatga 5281 tggcatgctt ctatattatt ttctaaaaga tttaaagttt tgccttctcc atttagactt 5341 ataattcact ggaatttttt tgtgtgtatg gtatgacata tgggttccct tttatttttt 5401 acatataaat atatttccct gtttttctaa aaaagaaaaa gatcatcatt ttcccattgt 5461 aaaatgccat atttttttca taggtcactt acatatatca atgggtctgt ttctgagctc 5521 tactctattt tatcagcctc actgtctatc cccacacatc tcatgctttg ctctaaatct 5581 tgatatttag tggaacattc tttcccattt tgttctacaa gaatattttt gttattgtct 5641 tttgggcttc tatatacatt ttagaatgag gttggcaagt tctcgacctg cagccaagct 5701 tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca 5761 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata 5821 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca 5881 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc 5941 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc 6001 aaggacctga aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt 6061 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg 6121 cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc 6181 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact 6241 acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg 6301 agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat 6361 actccaatac tcctgaaata gttcattatg gacagcgcag aagagctggg gagaattaat 6421 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 6481 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 6541 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 6601 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 6661 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6721 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6781 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6841 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6901 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6961 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 7021 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 7081 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7141 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7201 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7261 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7321 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 7381 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 7441 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 7501 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 7561 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7621 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7681 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7741 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7801 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 7861 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7921 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7981 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 8041 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 8101 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 8161 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 8221 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 8281 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 8341 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 8401 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 8461 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 8521 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 8581 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg 8641 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc 8701 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc 8761 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt 8821 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac 8881 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg 8941 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg 9001 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aagcagccca 9061 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg 9121 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca 9181 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag 9241 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag 9301 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc 9361 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc 9421 agaaaaaggg gggaa //