LOCUS Exported 7175 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pId1-IRES-YFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7175) AUTHORS Nam HS, Benezra R TITLE High levels of Id1 expression define B1 type adult neural stem cells. JOURNAL Cell Stem Cell. 2009 Nov 6. 5(5):515-26. PUBMED 19896442 REFERENCE 2 (bases 1 to 7175) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7175 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 1189..1211 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1203..1220 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1204..1220 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1227..1245 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1352..1369) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1579..1598 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 1759..2478 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1804..1825) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2065..2084) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2412..2433 /label=EGFP-C /note="EGFP, forward primer" promoter complement(2494..2512) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2495..2512) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2532..2549) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2538..2762 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2808..3236 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2895..2914) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3105..3126 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3245..3265) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3250..3579 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3430..3565 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3492..3511 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3646..4440 /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(3700..3719) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4310..4329 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4614..4735 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4651..4670) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4705..4724 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4784..4800) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4784..4800) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4797..4819) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4808..4824 /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(4832..4862) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4877..4898 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5015..5032) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5186..5774) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5266..5285) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5945..6805) /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 6568..6587 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6806..6910) /gene="bla" /label=AmpR promoter primer_bind complement(6985..7004) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt 661 ggtaccgagc tcggatccac catgaaggtc gccagtggca gtgccgcagc cgctgcaggc 721 cctagctgtt cgctgaaggc gggcaggaca gcgggcgagg tggtacttgg tctgtcggag 781 caaagcgtgg ccatctcgcg ctgcgctggg acgcgcctgc ccgccttgct ggacgagcag 841 caggtgaacg tcctgctcta cgacatgaac ggctgctact cacgcctcaa ggagctggtg 901 cccaccctgc cccagaaccg caaagtgagc aaggtggaga tcctgcagca tgtaatcgac 961 tacatcaggg acctgcagct ggagctgaac tcggagtctg aagtcgggac caccggaggc 1021 cggggactgc ctgtccgcgc cccgctcagc accctgaacg gcgagatcag tgccttggcg 1081 gccgaggcgg catgtgttcc agccgacgat cgcatcttgt gtcgctgaga attctggcga 1141 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac 1201 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttaattccgg 1261 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc 1321 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg 1381 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg 1441 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca 1501 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata 1561 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc 1621 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt 1681 gtttagtcga ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt 1741 gaaaaacacg atgataccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 1801 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 1861 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg 1921 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc 1981 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 2041 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 2101 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 2161 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc 2221 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac 2281 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 2341 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag 2401 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 2461 gacgagctgt acaagtaatc tagagggccc tattctatag tgtcacctaa atgctagagc 2521 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc 2581 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga 2641 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga 2701 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat 2761 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag 2821 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 2881 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 2941 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 3001 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 3061 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 3121 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc 3181 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 3241 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 3301 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 3361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 3421 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 3481 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag 3541 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata 3601 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 3661 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 3721 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 3781 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 3841 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3901 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3961 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 4021 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 4081 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 4141 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc 4201 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 4261 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 4321 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 4381 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 4441 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 4501 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 4561 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 4621 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 4681 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 4741 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 4801 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 4861 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 4921 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 4981 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 5041 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 5101 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 5161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 5221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 5281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 5341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 5401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 5461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 5521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 5581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 5641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 5701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 5761 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 5821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 5881 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 5941 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6001 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 6061 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6121 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 6181 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 6241 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 6301 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 6361 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 6421 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 6481 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 6541 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 6601 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 6661 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 6721 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 6781 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 6841 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 6901 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc 6961 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta 7021 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac 7081 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc 7141 gctgcttcgc gatgtacggg ccagatatac gcgtt //