LOCUS Exported 6738 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS GFP-PIPK1 gamma 90 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6738) AUTHORS Di Paolo G, Pellegrini L, Letinic K, Cestra G, Zoncu R, Voronov S, Chang S, Guo J, Wenk MR, De Camilli P TITLE Recruitment and regulation of phosphatidylinositol phosphate kinase type 1 gamma by the FERM domain of talin. JOURNAL Nature. 2002 Nov 7. 420(6911):85-9. PUBMED 12422219 REFERENCE 2 (bases 1 to 6738) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6738 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 16..319 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 320..523 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 474..494 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 562..571 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 568..1284 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 568..1284 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(613..634) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(874..893) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1221..1242 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3481..3602 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3518..3537) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3572..3591 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3609..4064) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3746..3767) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3958..3977 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4091..4195 /gene="bla" /label=AmpR promoter promoter 4197..4554 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4220..4240) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4405..4540 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4467..4486 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4589..5383 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4643..4662) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5253..5272 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5571..5590) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5615..5662 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5991..6579 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6480..6499 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca 541 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg 601 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 661 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 721 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 781 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 841 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 901 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 961 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc 1021 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac 1081 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac 1141 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac 1201 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact 1261 ctcggcatgg acgagctgta caagtccggc cggactcaga tctcgagctc aagcttcgaa 1321 ttcatggagc tggaggtacc ggacgaggcg gagagcgctg aggcgggggc cgtgccctcg 1381 gaggcggcgt gggcggcaga gagcggggcg gcggcaggtt tggctcagaa gaaggcggcc 1441 ccaacagagg ttctgtccat gacggcacag ccgggccctg gccatgggaa gaagttgggc 1501 catcgaggtg tggacgcatc cggcgaaacc acctacaaga agaccacctc ctccaccctg 1561 aagggtgcca tccagctggg catcggctac accgtgggcc acctgagctc caagcccgaa 1621 cgcgacgtgc tcatgcagga cttctacgtg gtggagagca tcttcttccc cagcgaaggc 1681 agcaacctca cccccgccca ccacttccag gacttccgct tcaagaccta tgcacctgtc 1741 gccttccgct acttccggga gctctttggg atccggccag atgattactt gtactccctg 1801 tgcaatgagc cgctgatcga gctgtccaac ccgggcgcca gtggctccct cttctacgtc 1861 accagcgacg acgagttcat catcaagacc gtcatgcaca aggaggccga gttcctgcag 1921 aagctgctcc ctggctacta catgaacctc aaccagaacc cgcggacgct gctgcccaag 1981 ttctatgggc tgtactgcgt gcagtcgggg ggcaagaaca tccgcgtcgt ggtcatgaac 2041 aacatcctgc cccgcgtggt caagatgcac ctcaagttcg acctcaaggg ctccacctac 2101 aagcggcgcg ccagcaagaa ggagaaggag aagagcttcc ccacctacaa ggacctggac 2161 ttcatgcagg acatgcccga ggggctcctg ctggacgccg acaccttcag cgccctggtc 2221 aagacgctgc agcgggactg cctggtcctg gaaagtttca agatcatgga ctacagcctg 2281 ctgctgggcg tgcacaacat cgaccagcac gagcgcgagc ggcaggcgca gggcgcccag 2341 agcacctcag atgagaagcg gcctgtgggc cagaaggcgc tctactccac ggccatggag 2401 tccatccagg gtggcgccgc gcgcggggag gccatcgaat cggatgacac gatgggcggg 2461 atccccgctg tgaacggccg cggggagcgg ctgctgctgc acattggcat catcgacatc 2521 ctgcagtcct acaggttcat caagaaactg gagcacacct ggaaggccct cgtccacgat 2581 ggggacacgg tgtccgtcca ccgccccagc ttctatgccg agcgcttttt caagttcatg 2641 agcaacacgg tctttcggaa gaactcctcc ctgaagtcct cgccctccaa gaaggggcgc 2701 ggcggagcct tgctagctgt gaaaccgctg gggcccaccg ctgccttctc ggccagccag 2761 atccctagcg agcgggagga ggcccagtac gacctgcggg gggcccgcag ctaccccacg 2821 ctggaggacg aaggccggcc cgacctcctg ccctgcacgc caccttcttt cgaagaagcc 2881 actacagcct ccattgccac gactctgtca tccacatccc tctccattcc tgagcggtcc 2941 ccctcggaga cgtcggagca gccgcggtac aggcggcgca cacagtcgtc tggacaggat 3001 ggcaggccgc aggaggagcc acccgcggaa gaggatctgc agcagattac agtgcaggtg 3061 gagcctgcgt gcagcgtgga gattgtggtc cccaaagagg aggacgcagg ggtggaggct 3121 tccccggccg gtgcctctgc tgctgttgaa gtagaaactg ccagccaggc ctcagacgag 3181 gagggcgcac ctgccagcca ggcctcggac gaggaggacg cgcccgccac cgacatctac 3241 tttcccaccg atgagaggag ctgggtgtac tccccgctcc actatagcgc ccaggccccc 3301 ccggcctccg acggcgagag cgacacataa gtcgacggta ccgcgggccc gggatccacc 3361 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta 3421 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 3481 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 3541 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 3601 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 3661 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 3721 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 3781 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 3841 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 3901 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 3961 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 4021 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg 4081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 4141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga 4201 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 4261 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 4321 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 4381 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 4441 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 4501 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga 4561 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 4621 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 4681 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 4741 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc 4801 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 4861 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 4921 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 4981 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 5041 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 5101 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 5161 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 5221 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 5281 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 5341 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 5401 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 5461 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 5521 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg 5581 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg 5641 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 5701 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 5761 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 5821 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt 5881 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 5941 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 6001 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 6061 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 6121 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 6181 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 6241 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 6301 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 6361 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 6421 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 6481 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 6541 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 6601 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 6661 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg 6721 gtcattagtt catagccc //