LOCUS Exported 8354 bp ds-DNA circular SYN 14-SEP-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HOXA9-MSCV-full-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8354) AUTHORS Shen WF, Hu YL, Uttarwar L, Passegue E, Largman C TITLE MicroRNA-126 regulates HOXA9 by binding to the homeobox. JOURNAL Mol Cell Biol. 2008 Jul . 28(14):4609-19. PUBMED 18474618 REFERENCE 2 (bases 1 to 8354) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 14, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8354 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..517 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 581..922 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 989..1405 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1335..1357 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1373..1389 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1417..1440 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter 3436..3935 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3471..3494) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 3868..3887 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 3942..3951 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3948..4667 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3948..4664 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3993..4014) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4254..4273) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4601..4622 /label=EGFP-C /note="EGFP, forward primer" LTR 4749..5263 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 5425..5441 /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(5449..5479) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5494..5515 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5632..5649) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5803..6391) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5883..5902) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6562..7422) /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 7185..7204 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7423..7527) /gene="bla" /label=AmpR promoter primer_bind 7595..7613 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7651..7673) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7773..7792 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7986..8008 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8136..8155 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc 61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag 121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa 181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt 241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt 301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac 361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa 421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct 481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga 541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt 601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct 661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg 721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc 781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg 841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg 901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac 961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac 1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa 1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg 1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg 1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga 1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc 1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc 1381 agccctcact ccttctctag gcgccggaat tctatggact acaaggacga cgatgacaag 1441 cttaagaggc ctcaggatcc caccaccggg gccctgggca actactatgt ggactccttc 1501 ctgctgggcg ccgacgctgc tgatgagctg ggtgcgggac gctacgctcc agggaccctg 1561 ggtcaacccc caaggcaggc ggcagctctg gccgaacacc ccgacttcag tccttgcagc 1621 ttccagtcca aggcggcggt gtttggtgcc tcgtggaacc cagtgcacgc ggcgggcgcc 1681 aatgcggtgc ctgctgcagt gtatcatcac caccaccacc cctacgtgca tccccaggcg 1741 cccgtggcgg cggcggcgcc ggacggcagg tatatgcgct cctggctgga acccacgccc 1801 ggtgcgctct ccttcgcggg cttaccctcc agccggcctt atggcattaa acctgaaccg 1861 ctctcggcca gaaggggtga ctgtcccacg cttgacactc acactttgtc cctgactgac 1921 tatgcttgtg gttctcctcc agttgataga gaaaaacaac ccagcgaagg cgccttctcc 1981 gaaaacaatg ccgagaatga gagcggcgga gacaagcccc ccatcgatcc caataacccg 2041 gctgccaact ggctacatgc tcgctccact cggaagaagc gatgccccta cacaaaacac 2101 cagacgctgg aactggagaa ggagtttctg tttaacatgt acctcacacg ggaccgcagg 2161 tacgaggtgg cccggctgct caacctcacc gaaaggcagg tcaagatctg gttccagaac 2221 cgcaggatga aaatgaagaa aatcaacaag gaccgagcaa aagacgagtg agccttttag 2281 gggcacattt aaaaagagag caagctagaa agaaaaagaa aggactgtcc gtctccctct 2341 gtctcctctc ccccaaaccc agcctccacc cgcacaaagg ggctctaaat cccaggcctc 2401 atctccccac tggcagcccg tgctcaggct ggctcttagg cctgcggctt tgatggagga 2461 ggtattgtaa gctttccatt ttatagaagg cacacacaca cacaagggag ggcattagcg 2521 ctattggctg tatgtgctag cttgtatata tatatatata tttaaaaaaa atctacctgc 2581 ttctgacttt aagcaaaagg aaagaaaact acctttttat ataatgcaca actgttgatg 2641 actggctgta tagtttttag tctctgtagc taatttaatt tgctcttcgt gtggcagatc 2701 attctgccaa aatacttgaa cactgtgttt tattgtggtg attatgtttt gtgactcaaa 2761 cttctgtgct gggtgaagta cccattgtga ttgtggaagc tagaattcaa tttgaactca 2821 ggttgtttat ggggggagag aagctgtggc ataatgtata gctctgtagt cttctgtgta 2881 gctagactgt taacctacga ttgtaaacta gctgtaagga tttcccagtg aaatcagatt 2941 tggaaagaaa agagagatag ttctccggga tgcatagatt catgatttat ctacttttga 3001 aatgtgggct tcttcatctt tacgttctag actccaattt tgtccaatgt atatagaatc 3061 tccatattaa agaaatattc ataatcagaa agcttgaata ttgtttttgc accagaggag 3121 aagtgaggaa attcggagct atacgtgtgt gcagagggtt gctacctatg ggtttacgct 3181 taattttaat tgggagaaag tgagtgctta ttgtaatgga gttaatttta ttgatgtacc 3241 ttatacacca tgaaactgct cttgggctac tgtagatttg tatccttggt gaatttgggg 3301 ttcccatcgg attgaacata cactgtatat tttgcaatag ttacctcaag gcctactgac 3361 caaattgttg tgtcgagata tttaactttt gccaaacgga atatattgat tcttctcgag 3421 agatctaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt 3481 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca 3541 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta 3601 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac 3661 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga 3721 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc 3781 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg 3841 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg 3901 cgctgttctc ctcttcctca tctccgggcc tttcggctgc agccaccatg gtgagcaagg 3961 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg 4021 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc 4081 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc 4141 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct 4201 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg 4261 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg 4321 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca 4381 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga 4441 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc 4501 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc 4561 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg 4621 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccaactcg 4681 acgttattcc cttcgaaggc ctatcgataa aataaaagat tttatttagt ctccagaaaa 4741 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt 4801 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga 4861 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 4921 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat 4981 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc 5041 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc 5101 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg 5161 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg 5221 tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga 5281 agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag 5341 ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag 5401 aaagagctgg ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc 5461 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 5521 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 5581 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 5641 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 5701 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 5761 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 5821 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 5881 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 5941 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6001 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6061 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6241 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 6421 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 6481 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 6541 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 6601 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 6661 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 6721 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 6781 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 6841 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 6901 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 6961 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7021 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7081 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7141 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7201 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7261 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7321 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7381 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 7441 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 7501 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 7561 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 7621 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 7681 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 7741 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 7801 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 7861 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 7921 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 7981 gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg 8041 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc 8101 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca 8161 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta 8221 gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat 8281 caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc 8341 cagaaaaagg gggg //