LOCUS Exported 9091 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS TagRFP-T-EEA1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9091) AUTHORS Navaroli DM, Bellve KD, Standley C, Lifshitz LM, Cardia J, Lambright D, Leonard D, Fogarty KE, Corvera S TITLE Rabenosyn-5 defines the fate of the transferrin receptor following clathrin-mediated endocytosis. JOURNAL Proc Natl Acad Sci U S A. 2012 Feb 21;109(8):E471-80. Epub 2012 Jan 30. PUBMED 22308388 REFERENCE 2 (bases 1 to 9091) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9091 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 44..347 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 348..551 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 502..522 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 590..599 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 596..1327 /codon_start=1 /product="photostable variant of TagRFP (Shaner et al., 2008)" /label=TagRFP-T /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV ARYCDLPSKLGHKLNGMDELYK" CDS 1337..5572 /codon_start=1 /product="human early endosome antigen 1" /label=EEA1 /translation="MLRRILQRTPGRVGSQGSDLDSSATPINTVDVNNESSSEGFICPQ CMKSLGSADELFKHYEAVHDAGNDSGHGGESNLALKRDDVTLLRQEVQDLQASLKEEKW YSEELKKELEKYQGLQQQEAKPDGLVTDSSAELQSLEQQLEEAQTENFNIKQMKDLFEQ KAAQLATEIADIKSKYDEERSLREAAEQKVTRLTEELNKEATVIQDLKTELLQRPGIED VAVLKKELVQVQTLMDNMTLERERESEKLKDECKKLQSQYASSEATISQLRSELAKGPQ EVAVYVQELQKLKSSVNELTQKNQTLTENLLKKEQDYTKLEEKHNEESVSKKNIQATLH QKDLDCQQLQSRLSASETSLHRIHVELSEKGEATQKLKEELSEVETKYQHLKAEFKQLQ QQREEKEQHGLQLQSEINQLHSKLLETERQLGEAHGRLKEQRQLSSEKLMDKEQQVADL QLKLSRLEEQLKEKVTNSTELQHQLDKTKQQHQEQQALQQSTTAKLREAQNDLEQVLRQ IGDKDQKIQNLEALLQKSKENISLLEKEREDLYAKIQAGEGETAVLNQLQEKNHTLQEQ VTQLTEKLKNQSESHKQAQENLHDQVQEQKAHLRAAQDRVLSLETSVNELNSQLNESKE KVSQLDIQIKAKTELLLSAEAAKTAQRADLQNHLDTAQNALQDKQQELNKITTQLDQVT AKLQDKQEHCSQLESHLKEYKEKYLSLEQKTEELEGQIKKLEADSLEVKASKEQALQDL QQQRQLNTDLELRATELSKQLEMEKEIVSSTRLDLQKKSEALESIKQKLTKQEEEKQIL KQDFETLSQETKIQHEELNNRIQTTVTELQKVKMEKEALMTELSTVKDKLSKVSDSLKN SKSEFEKENQKGKAAILDLEKTCKELKHQLQVQMENTLKEQKELKKSLEKEKEASHQLK LELNSMQEQLIQAQNTLKQNEKEEQQLQGNINELKQSSEQKKKQIEALQGELKIAVLQK TELENKLQQQLTQAAQELAAEKEKISVLQNNYEKSQETFKQLQSDFYGRESELLATRQD LKSVEEKLSLAQEDLISNRNQIGNQNKLIQELKTAKATLEQDSAKKEQQLQERCKALQD IQKEKSLKEKELVNEKSKLAEIEEIKCRQEKEITKLNEELKSHKLESIKEITNLKDAKQ LLIQQKLELQGKADSLKAAVEQEKRNQQILKDQVKKEEEELKKEFIEKEAKLHSEIKEK EVGMKKHEENEAKLTMQITALNENLGTVKKEWQSSQRRVSELEKQTDDLRGEIAVLEAT VQNNQDERRALLERCLKGEGEIEKLQTKVLELQRKLDNTTAAVQELGRENQSLQIKHTQ ALNRKWAEDNEVQNCMACGKGFSVTVRRHHCRQCGNIFCAECSAKNALTPSSKKPVRVC DACFNDLQG" polyA_signal 5862..5983 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5899..5918) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5953..5972 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5990..6445) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6127..6148) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6339..6358 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6472..6576 /gene="bla" /label=AmpR promoter promoter 6578..6935 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6601..6621) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6786..6921 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6848..6867 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6970..7764 /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(7024..7043) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7634..7653 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7952..7971) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7996..8043 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 8372..8960 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8861..8880 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 61 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 541 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt 601 gtctaagggc gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac 661 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac 721 ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct 781 ggctaccagc ttcatgtacg gcagcagaac cttcatcaac cacacccagg gcatccccga 841 tttctttaag cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga 901 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa 961 cgtcaagatc agaggggtga acttcccatc caacggccct gtgatgcaga agaaaacact 1021 cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctgg aaggcagaac 1081 cgacatggcc ctgaagctcg tgggcggggg ccacctgatc tgcaacttca agaccacata 1141 cagatccaag aaacccgcta agaacctcaa gatgcccggc gtctactatg tggaccacag 1201 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg aggtggctgt 1261 ggccagatac tgcgacctcc ctagcaaact ggggcacaaa cttaatggca tggacgagct 1321 gtacaagact cgagaaatgt taaggaggat tttacagagg actcctggga gagttggctc 1381 tcaaggttct gatttagatt catcagcaac tcctataaac acagtggacg tcaataatga 1441 aagctcttca gaaggtttca tatgtcccca gtgtatgaaa tctcttggat ctgctgatga 1501 acttttcaaa cattatgaag ctgttcatga tgctggtaat gactcaggtc atggaggaga 1561 gtctaatctt gctttgaagc gagatgatgt aacactgctc agacaagagg tccaagacct 1621 acaggcttca cttaaggaag aaaaatggta ctcggaagaa ttaaagaagg aattagaaaa 1681 atatcaaggg ctgcagcagc aagaggccaa acctgatggg ttggtgactg attcatcagc 1741 agaactacag tctttggaac agcaattaga agaagcccaa acagaaaatt ttaatattaa 1801 gcaaatgaaa gacttatttg aacagaaagc agcccaactt gctactgaaa ttgcagatat 1861 aaagtcaaag tatgatgaag aaaggagtct tcgagaagct gctgaacaaa aagtgacacg 1921 tctgacagaa gaattaaaca aagaggcaac tgtaattcaa gatctgaaga cggaactgct 1981 tcagagacct ggtatagaag atgttgccgt gctaaagaaa gaactggtcc aagttcaaac 2041 actaatggat aacatgacct tggaacgtga gcgagaatct gaaaaactca aagatgaatg 2101 caaaaaattg cagtcacaat atgctagctc agaggccaca ataagccagc taaggagtga 2161 acttgccaaa ggcccccagg aagttgctgt atatgtacag gaactacaaa aactgaaaag 2221 ttcagttaat gaattaacac aaaaaaatca gaccttgaca gaaaacttgc tgaaaaaaga 2281 acaagactat actaagttag aggagaaaca taatgaagaa tctgtgagta aaaagaatat 2341 tcaggcaacc cttcatcaaa aagacctaga ttgtcaacag cttcagtcaa gattgtctgc 2401 atctgaaacc tcactgcata gaatacatgt agaactaagt gaaaaaggag aagctactca 2461 aaagctcaaa gaagaattat ctgaggtaga gaccaagtac cagcatctaa aggcggagtt 2521 taagcagcta caacaacaga gagaagaaaa ggagcagcat gggttacaac tccaaagtga 2581 aattaatcaa ttacatagca aacttctaga gacagagcgc caactagggg aagctcatgg 2641 taggctgaag gaacagagac agctttcaag tgaaaagttg atggataaag aacaacaagt 2701 ggctgattta caactcaaac tttctcggtt agaagagcag ttgaaggaaa aagttacaaa 2761 ttctacagaa ttgcagcatc aattagataa aacaaagcaa cagcatcaag aacaacaggc 2821 tcttcagcaa agcaccacgg caaaacttcg agaagctcag aatgatttgg aacaagttct 2881 acgtcaaatt ggcgataagg accaaaagat ccagaacctt gaagctttat tacagaagag 2941 taaagaaaat atttcattac tagaaaaaga aagagaagat ctttatgcaa aaattcaggc 3001 tggtgaagga gagactgctg ttcttaacca gttacaagaa aaaaaccata cactacagga 3061 gcaagtaact caactaacag agaagctgaa gaatcagtca gaaagtcata aacaagccca 3121 ggagaatttg catgaccagg tacaagagca gaaggcacat cttagagctg cacaagaccg 3181 tgtcctttcc ctagaaacta gtgtcaatga attaaatagt caattaaatg aaagcaagga 3241 gaaggtctcc cagcttgaca tacagattaa agccaaaacc gaactattac tatcagcaga 3301 agcagcaaaa actgctcaaa gagctgatct tcagaatcat ttggacacag ctcaaaatgc 3361 attacaagat aaacagcagg agttaaataa gattactact cagttggatc aggtcactgc 3421 aaagttacaa gacaagcaag aacattgcag tcagctggaa agtcatctta aagaatataa 3481 agagaaatac ctctctttag aacagaaaac cgaagagcta gaaggtcaaa ttaagaaact 3541 agaagctgat agtcttgaag ttaaagcaag caaggagcag gctttgcaag atctacaaca 3601 gcaaagacag ctgaacacag atttagagct cagagccaca gaattgagta aacaacttga 3661 aatggagaag gaaatagtat ccagtacaag attggatcta cagaaaaaat ctgaagccct 3721 tgaaagtatc aagcaaaagc ttaccaagca agaggaagaa aaacaaatcc tgaaacaaga 3781 ttttgaaact ttaagtcaag aaacaaagat tcagcatgag gaattgaata acagaattca 3841 aacaacagta acagaactac aaaaagtgaa aatggagaaa gaagctttaa tgacagagct 3901 ttctacagta aaggacaaac tatcaaaagt ttctgattct ttgaaaaact ctaaaagtga 3961 atttgaaaag gagaatcaga aaggaaaagc cgctatatta gacttggaaa aaacttgcaa 4021 agaattaaag catcaacttc aagtgcagat ggaaaacaca cttaaggaac agaaggaact 4081 gaaaaagtca cttgaaaaag agaaggaggc ttctcatcag ttgaaattgg aactcaattc 4141 aatgcaggaa caacttatac aggcccagaa tactttaaaa caaaatgaaa aagaagagca 4201 acaacttcag gggaacataa atgagctaaa gcaatcaagt gaacagaaga aaaaacaaat 4261 tgaagcactc caaggagagc ttaaaattgc tgttttacag aagacagagc ttgagaataa 4321 actacagcag cagttaacac aggcagccca ggaacttgca gcagagaaag agaaaatatc 4381 agtattacaa aacaactatg aaaaaagtca ggaaactttc aaacagcttc aatctgattt 4441 ctatgggagg gaatctgaac ttctagccac caggcaagat cttaagtctg tagaagagaa 4501 gctttctcta gcacaggagg acttgatttc aaacagaaat caaattggaa atcaaaataa 4561 attgattcaa gaactgaaga ctgccaaggc tacattggag caggattcag caaagaaaga 4621 acagcaattg caggagcgat gtaaagcact acaagacatt cagaaagaaa agtcactgaa 4681 agaaaaagaa ctggtaaatg agaagtctaa attggcagag atagaagaaa ttaaatgtag 4741 acaagaaaaa gaaatcacta aactaaacga agaactcaag tcccacaaac tagaaagcat 4801 aaaggagata acaaatctta aagatgctaa acagcttcta attcagcaga aattagaact 4861 tcaaggaaaa gcggactccc tgaaggcagc tgttgaacag gagaagagaa atcagcagat 4921 actaaaagac caggtgaaaa aggaagaaga ggagctgaag aaagaattta ttgagaaaga 4981 agctaagttg cattccgaaa taaaagaaaa ggaagtagga atgaagaagc atgaagaaaa 5041 tgaggctaaa cttaccatgc agattacagc attaaatgaa aacttaggca ctgtgaagaa 5101 ggagtggcaa tctagtcaac ggagagttag tgagcttgag aaacaaacgg atgacttacg 5161 gggtgaaatt gcagtattag aagcaacggt tcagaataat caagatgaaa ggagagcact 5221 actggaaaga tgtcttaaag gagaaggtga aatagaaaag cttcaaacca aagtattaga 5281 attgcaaaga aagctggata atacaactgc agcagtgcag gagctgggca gagaaaacca 5341 atcacttcag atcaaacata cacaagcgtt gaatagaaag tgggccgaag acaatgaagt 5401 acaaaactgt atggcctgtg ggaaaggctt ttcagtaaca gtgagacggc atcactgccg 5461 acagtgtgga aatatcttct gtgctgaatg ttcagccaaa aatgccttaa ctccttcctc 5521 caagaagcct gttcgtgtct gtgatgcatg tttcaatgac ttgcaaggat aatgggttat 5581 cacaacttca gagtaatatt acactaacat tagattttta ataaatgtac ttaatagagg 5641 tcttggacta ctatttggtt tggacactgg ttgcaatact agaccaaata ggaattagta 5701 tattttggaa tgttatggat atcgcggccg ctggatccac cggatctaga taactgatca 5761 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 5821 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt 5881 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5941 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc 6001 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 6061 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 6121 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 6181 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 6241 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 6301 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 6361 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 6421 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 6481 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 6541 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt 6601 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 6661 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 6721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 6781 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6841 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6901 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga 6961 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 7021 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 7081 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 7141 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 7201 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 7261 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 7321 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 7381 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 7441 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc 7501 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 7561 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 7621 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 7681 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 7741 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 7801 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 7861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 7921 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 7981 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg 8041 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc 8101 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg 8161 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact 8221 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 8281 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 8341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 8401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 8461 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 8521 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 8581 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 8641 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 8701 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 8761 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 8821 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 8881 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 8941 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 9001 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 9061 ttaccgccat gcattagtta ttaatagtaa t //