LOCUS Exported 5365 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS GFP-HP1gamma SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5365) AUTHORS Cheutin T, McNairn AJ, Jenuwein T, Gilbert DM, Singh PB, Misteli T TITLE Maintenance of stable heterochromatin domains by dynamic HP1 binding. JOURNAL Science. 2003 Jan 31. 299(5607):721-5. PUBMED 12560555 REFERENCE 2 (bases 1 to 5365) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5365 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 77..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" primer_bind 581..605 /label=LNCX /note="Human CMV promoter, forward primer" promoter 815..833 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 815..832 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 846..865 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 918..927 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 930..947 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 948..1658 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELY" primer_bind complement(978..1006) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1601..1621 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" polyA_signal 2196..2330 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(2366..2384) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2405..2421) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2405..2421) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2418..2440) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2429..2445 /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(2453..2483) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2498..2519 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2636..2653) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2807..3395) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2887..2906) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3566..4426) /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 4189..4208 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4427..4531) /gene="bla" /label=AmpR promoter rep_origin complement(4557..5012) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4694..4715) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4906..4925 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5139..5161 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5153..5170 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5154..5170 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 5177..5196 /label=T7 /note="T7 promoter, forward primer" promoter 5177..5195 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 5205..5339 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" ORIGIN 1 acgtcgcatg ctcctctagt 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 agctcgttta gtgaaccgtc 601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat 661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg 721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca 781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat 841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctgg 901 gccgctctag aactagcgcc accatgggac atcaccatca ccatcacatg agtaaaggag 961 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc 1021 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta 1081 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttca 1141 cttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat gactttttca 1201 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga 1261 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt 1321 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact 1381 ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact 1441 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa 1501 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat 1561 ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa 1621 cagctgctgg gattacacat ggcatggatg aactatacaa gggatccgga aaaaaacaga 1681 atggaaagag taaaaaagtt gaagaggcag agcctgaaga atttgtcgtg gaaaaagtac 1741 tagatcgacg tgtagtgaat gggaaagtgg aatatttcct gaagtggaag ggatttacag 1801 atgctgacaa tacttgggaa cctgaagaaa atttagattg tccagaattg attgaagcgt 1861 ttcttaactc tcagaaagct ggcaaagaaa aagatggtac aaaaagaaaa tctttatctg 1921 acagtgaatc tgatgacagc aaatcaaaga agaaaagaga tgctgctgac aaaccaagag 1981 gatttgccag aggtcttgat cctgaaagaa taattggtgc cacagacagc agtggagaat 2041 tgatgtttct catgaaatgg aaagattcag atgaggcaga cttggtgctg gcgaaagagg 2101 caaatatgaa gtgtcctcaa attgtaattg ctttttatga agagagacta acttggcatt 2161 cttgtccaga agatgaagct caataagctt aacccaactt gtttattgca gcttataatg 2221 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 2281 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gacctcgagg 2341 gggggcccgg taccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat 2401 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 2461 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 2521 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 2581 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 2641 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 2701 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 2761 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 2821 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 2881 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 2941 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 3001 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 3061 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 3121 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 3181 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 3241 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 3301 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 3361 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 3421 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 3481 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 3541 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 3601 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 3661 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 3721 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 3781 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 3841 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 3901 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 3961 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 4021 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 4081 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 4141 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 4201 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 4261 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 4321 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 4381 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 4441 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 4501 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat 4561 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 4621 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 4681 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 4741 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc 4801 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 4861 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 4921 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 4981 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg 5041 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg 5101 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 5161 gacggccagt gaattgtaat acgactcact atagggcgaa ttggaacttg tttattgcag 5221 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5281 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg 5341 atccccccac cgcggtggcg gcccg //