LOCUS Exported 5804 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes for human CXCR4 coding sequence tagged with the LSS-mOrange FP. ACCESSION . VERSION . KEYWORDS hCXCR4-LSSmOrange SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5804) TITLE Padilla Parra unpublished receptor-FP plasmids REFERENCE 2 (bases 1 to 5804) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5804 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 135..438 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 439..642 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 593..613 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 1425..1436 /label=ZF binding site /bound_moiety="CCR5TC zinc-finger domain" /note="target sequence for the CCR5TC zinc-finger domain from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 2008; Gross et al., 2013)" regulatory 1767..1776 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1773..2480 /codon_start=1 /product="monomeric orange fluorescent protein with a large Stokes shift (Shcherbakova et al., 2012)" /label=LSSmOrange /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind 2186..2205 /label=mCherry-F /note="mCherry, forward primer" misc_feature 2486..2542 /label=MCS /note="multiple cloning site" polyA_signal 2666..2787 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2703..2722) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2757..2776 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2794..3249) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2931..2952) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3143..3162 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3276..3380 /gene="bla" /label=AmpR promoter promoter 3382..3739 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3405..3425) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3590..3725 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3652..3671 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3774..4568 /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(3828..3847) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4438..4457 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4756..4775) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4800..4847 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5176..5764 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5665..5684 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 61 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca 121 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 181 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact 241 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 301 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 361 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 421 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg 481 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg 541 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 601 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag 661 atccgctagc gctaccggac tcagatctcg agctcaagct tatggagggg atcagtatat 721 acacttcaga taactacacc gaggaaatgg gctcagggga ctatgactcc atgaaggaac 781 cctgtttccg tgaagaaaat gctaatttca ataaaatctt cctgcccacc atctactcca 841 tcatcttctt aactggcatt gtgggcaatg gattggtcat cctggtcatg ggttaccaga 901 agaaactgag aagcatgacg gacaagtaca ggctgcacct gtcagtggcc gacctcctct 961 ttgtcatcac gcttcccttc tgggcagttg atgccgtggc aaactggtac tttgggaact 1021 tcctatgcaa ggcagtccat gtcatctaca cagtcaacct ctacagcagt gtcctcatcc 1081 tggccttcat cagtctggac cgctacctgg ccatcgtcca cgccaccaac agtcagaggc 1141 caaggaagct gttggctgaa aaggtggtct atgttggcgt ctggatccct gccctcctgc 1201 tgactattcc cgacttcatc tttgccaacg tcagtgaggc agatgacaga tatatctgtg 1261 accgcttcta ccccaatgac ttgtgggtgg ttgtgttcca gtttcagcac atcatggttg 1321 gccttatcct gcctggtatt gtcatcctgt cctgctattg cattatcatc tccaagctgt 1381 cacactccaa gggccaccag aagcgcaagg ccctcaagac cacagtcatc ctcatcctgg 1441 ctttcttcgc ctgttggctg ccttactaca ttgggatcag catcgactcc ttcatcctcc 1501 tggaaatcat caagcaaggg tgtgagtttg agaacactgt gcacaagtgg atttccatca 1561 ccgaggccct agctttcttc cactgttgtc tgaaccccat cctctatgct ttccttggag 1621 ccaaatttaa aacctctgcc cagcacgcac tcacctctgt gagcagaggg tccagcctca 1681 agatcctctc caaaggaaag cgaggtggac attcatctgt ttccaccgag tctgagtctt 1741 caagttttca ctccagcgca ccggtcgcca ccatggtgag caagggcgag gagaataaca 1801 tggccatcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc 1861 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcttt cagaccgtta 1921 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcttg tcccctcagt 1981 tcacctacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacctcaagc 2041 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg 2101 tgaccgtgac tcaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc 2161 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggcatggagg 2221 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgaggac aagctcaggc 2281 tgaagctgaa ggacggcggc cactacacct ccgaggtcaa gaccacctac aaggccaaga 2341 agcccgtgca gttgcccggc gcctacatcg tcgacatcaa gttggacatc acctcccaca 2401 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg 2461 gcatggacga gctgtacaag tactcagatc tcgagctcaa gcttcgaatt ctgcagtcga 2521 cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca gccataccac 2581 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca 2641 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata 2701 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 2761 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa 2821 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 2881 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 2941 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 3001 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc 3061 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 3121 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 3181 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac 3241 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 3301 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 3361 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt 3421 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 3481 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 3541 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 3601 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 3661 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 3721 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg 3781 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3841 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3901 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg 3961 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 4021 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 4081 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 4141 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 4201 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 4261 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg 4321 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 4381 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4441 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4501 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4561 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 4621 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 4681 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 4741 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac 4801 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt 4861 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc 4921 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc 4981 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat 5041 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 5101 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 5161 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 5221 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 5281 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 5341 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 5401 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 5461 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 5521 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 5581 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 5641 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 5701 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 5761 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc tttt //