LOCUS Exported 5808 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes for human CXCR4 coding sequence tagged with the mTFP1 FP. ACCESSION . VERSION . KEYWORDS hCXCR4-mTFP1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5808) TITLE Padilla Parra unpublished receptor-FP plasmids REFERENCE 2 (bases 1 to 5808) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5808 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 90..393 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 394..597 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 548..568 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 1380..1391 /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 1722..1731 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1728..2435 /codon_start=1 /product="monomeric teal (cyan) fluorescent protein (Ai et al., 2006)" /label=mTFP1 /note="mammalian codon-optimized" /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA VARNSTDGMDELYK" misc_feature 2436..2501 /label=MCS /note="multiple cloning site" polyA_signal 2625..2746 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2662..2681) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2716..2735 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2753..3208) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2890..2911) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3102..3121 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3235..3339 /gene="bla" /label=AmpR promoter promoter 3341..3698 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3364..3384) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3549..3684 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3611..3630 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3733..4527 /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(3787..3806) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4397..4416 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4715..4734) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4759..4806 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5135..5723 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5624..5643 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca 61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt 601 ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttatgg 661 aggggatcag tatatacact tcagataact acaccgagga aatgggctca ggggactatg 721 actccatgaa ggaaccctgt ttccgtgaag aaaatgctaa tttcaataaa atcttcctgc 781 ccaccatcta ctccatcatc ttcttaactg gcattgtggg caatggattg gtcatcctgg 841 tcatgggtta ccagaagaaa ctgagaagca tgacggacaa gtacaggctg cacctgtcag 901 tggccgacct cctctttgtc atcacgcttc ccttctgggc agttgatgcc gtggcaaact 961 ggtactttgg gaacttccta tgcaaggcag tccatgtcat ctacacagtc aacctctaca 1021 gcagtgtcct catcctggcc ttcatcagtc tggaccgcta cctggccatc gtccacgcca 1081 ccaacagtca gaggccaagg aagctgttgg ctgaaaaggt ggtctatgtt ggcgtctgga 1141 tccctgccct cctgctgact attcccgact tcatctttgc caacgtcagt gaggcagatg 1201 acagatatat ctgtgaccgc ttctacccca atgacttgtg ggtggttgtg ttccagtttc 1261 agcacatcat ggttggcctt atcctgcctg gtattgtcat cctgtcctgc tattgcatta 1321 tcatctccaa gctgtcacac tccaagggcc accagaagcg caaggccctc aagaccacag 1381 tcatcctcat cctggctttc ttcgcctgtt ggctgcctta ctacattggg atcagcatcg 1441 actccttcat cctcctggaa atcatcaagc aagggtgtga gtttgagaac actgtgcaca 1501 agtggatttc catcaccgag gccctagctt tcttccactg ttgtctgaac cccatcctct 1561 atgctttcct tggagccaaa tttaaaacct ctgcccagca cgcactcacc tctgtgagca 1621 gagggtccag cctcaagatc ctctccaaag gaaagcgagg tggacattca tctgtttcca 1681 ccgagtctga gtcttcaagt tttcactcca gcgcaccggt cgccaccatg gtgagcaagg 1741 gcgaggagac cacaatgggc gtaatcaagc ccgacatgaa gatcaagctg aagatggagg 1801 gcaacgtgaa tggccacgcc ttcgtgatcg agggcgaggg cgagggcaag ccctacgacg 1861 gcaccaacac catcaacctg gaggtgaagg agggagcccc cctgcccttc tcctacgaca 1921 ttctgaccac cgcgttcgcc tacggcaaca gggccttcac caagtacccc gacgacatcc 1981 ccaactactt caagcagtcc ttccccgagg gctactcttg ggagcgcacc atgaccttcg 2041 aggacaaggg catcgtgaag gtgaagtccg acatctccat ggaggaggac tccttcatct 2101 acgagataca cctcaagggc gagaacttcc cccccaacgg ccccgtgatg cagaagaaga 2161 ccaccggctg ggacgcctcc accgagagga tgtacgtgcg cgacggcgtg ctgaagggcg 2221 acgtcaagca caagctgctg ctggagggcg gcggccacca ccgcgttgac ttcaagacca 2281 tctacagggc caagaaggcg gtgaagctgc ccgactatca ctttgtggac caccgcatcg 2341 agatcctgaa ccacgacaag gactacaaca aggtgaccgt ttacgagagc gccgtggccc 2401 gcaattccac cgacggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag 2461 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga 2521 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc 2581 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag 2641 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 2701 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta 2761 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac 2821 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg 2881 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa 2941 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt 3001 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt 3061 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga 3121 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc 3181 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga 3241 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 3301 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc 3361 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3421 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3481 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3541 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3601 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt 3661 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga 3721 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 3781 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 3841 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 3901 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 3961 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 4021 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 4081 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 4141 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 4201 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 4261 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 4321 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 4381 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 4441 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 4501 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 4561 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 4621 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 4681 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg 4741 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt 4801 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac 4861 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg 4921 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt 4981 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 5041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 5101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 5161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 5221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 5281 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtatca ccgcctacat 5341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 5401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 5461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 5521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 5581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 5641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 5701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 5761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgatt //