LOCUS Exported 6492 bp ds-DNA circular SYN 16-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMIG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6492) TITLE Viral vectors. REFERENCE 2 (bases 1 to 6492) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6492 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 123..842 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(168..189) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(429..448) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 776..797 /label=EGFP-C /note="EGFP, forward primer" LTR 992..1506 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 1668..1684 /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(1692..1722) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1737..1758 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1875..1892) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2046..2634) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2126..2145) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2805..3665) /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 3428..3447 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3666..3770) /gene="bla" /label=AmpR promoter primer_bind 3838..3856 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3894..3916) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4016..4035 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4229..4251 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4379..4398 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 4598..5114 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 5178..5519 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 5586..6002 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 5932..5954 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 5970..5986 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 6051..111 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(6195..6212) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 6422..6441 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" ORIGIN 1 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg 61 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa 121 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 181 acggcgacgt aaacggccac aagttcagcg tgtctggcga gggcgagggc gatgccacct 241 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 301 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 361 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 421 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 481 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 541 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 601 acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 661 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 721 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 781 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt 841 aatgaattaa ttaagaatta tcaccgcttc tattcagcca gtaaggcctg tctttaatgg 901 cctccggcat gagacacttt ctagagtcga cctgcagcca agcttatcga taaaataaaa 961 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa 1021 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag 1081 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg 1141 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc 1201 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac 1261 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg 1321 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag 1381 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt 1441 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc 1501 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata 1561 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa 1621 atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc 1681 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 1741 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 1801 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 1861 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 1921 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 1981 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 2041 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 2101 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 2161 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 2221 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 2281 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 2341 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 2401 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 2461 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 2521 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 2581 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 2641 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 2701 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 2761 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 2821 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 2881 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 2941 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 3001 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 3061 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 3121 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 3181 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 3241 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 3301 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 3361 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 3421 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 3481 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 3541 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 3601 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 3661 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 3721 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 3781 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 3841 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 3901 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 3961 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 4021 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta 4081 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg 4141 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg 4201 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa 4261 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca 4321 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg 4381 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga 4441 tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg 4501 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa 4561 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt 4621 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat 4681 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata 4741 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg 4801 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg 4861 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc 4921 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct 4981 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc 5041 cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc 5101 gggggtcttt catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga 5161 cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc 5221 gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct 5281 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga 5341 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt 5401 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc 5461 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc 5521 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa 5581 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg 5641 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct 5701 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc 5761 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg 5821 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct 5881 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac 5941 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg 6001 ccggaattag atctctcgag gttaacgaat tccgcccccc ccccccccta acgttactgg 6061 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 6121 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 6181 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 6241 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 6301 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 6361 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 6421 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 6481 tatgggatct ga //