LOCUS       Exported                6415 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression plasmid for untagged human CXCR4.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3-CXCR4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  AUTHORS   Heredia JD, Park J, Brubaker RJ, Szymanski SK, Gill KS, Procko E
  TITLE     Mapping Interaction Sites on Human Chemokine Receptors by Deep 
            Mutational Scanning.
  JOURNAL   J Immunol. 2018 Apr 20. pii: jimmunol.1800343. doi: 
            10.4049/jimmunol.1800343.
  PUBMED    29678950
REFERENCE   2  (bases 1 to 6415)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6415
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      1..10
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    730..741
                     /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)"
     primer_bind     complement(1106..1123)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1112..1336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1382..1810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1469..1488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1679..1700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1819..1839)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1824..2153
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2004..2139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2066..2085
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2220..3014
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2274..2293)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2884..2903
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3188..3309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3225..3244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3279..3298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3358..3374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3358..3374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3371..3393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3382..3398
                     /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(3406..3436)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3451..3472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3589..3606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3760..4348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3840..3859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4519..5379)
                     /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     5142..5161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5380..5484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5559..5578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5750..6129
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6130..6333
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6284..6304
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6378..6397
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6378..6396
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gccaccatgg aggggatcag tatatacact tcagataact acaccgagga aatgggctca
       61 ggggactatg actccatgaa ggaaccctgt ttccgtgaag aaaatgctaa tttcaataaa
      121 atcttcctgc ccaccatcta ctccatcatc ttcttaactg gcattgtggg caatggattg
      181 gtcatcctgg tcatgggtta ccagaagaaa ctgagaagca tgacggacaa gtacaggctg
      241 cacctgtcag tggccgacct cctctttgtc atcacgcttc ccttctgggc agttgatgcc
      301 gtggcaaact ggtactttgg gaacttccta tgcaaggcag tccatgtcat ctacacagtc
      361 aacctctaca gcagtgtcct catcctggcc ttcatcagtc tggaccgcta cctggccatc
      421 gtccacgcca ccaacagtca gaggccaagg aagctgttgg ctgaaaaggt ggtctatgtt
      481 ggcgtctgga tccctgccct cctgctgact attcccgact tcatctttgc caacgtcagt
      541 gaggcagatg acagatatat ctgtgaccgc ttctacccca atgacttgtg ggtggttgtg
      601 ttccagtttc agcacatcat ggttggcctt atcctgcctg gtattgtcat cctgtcctgc
      661 tattgcatta tcatctccaa gctgtcacac tccaagggcc accagaagcg caaggccctc
      721 aagaccacag tcatcctcat cctggctttc ttcgcctgtt ggctgcctta ctacattggg
      781 atcagcatcg actccttcat cctcctggaa atcatcaagc aagggtgtga gtttgagaac
      841 actgtgcaca agtggatttc catcaccgag gccctagctt tcttccactg ttgtctgaac
      901 cccatcctct atgctttcct tggagccaaa tttaaaacct ctgcccagca cgcactcacc
      961 tctgtgagca gagggtccag cctcaagatc ctctccaaag gaaagcgagg tggacattca
     1021 tctgtttcca ctgagtctga gtcttcaagt tttcactcca gctaatagct cgagtctaga
     1081 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1141 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1201 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1261 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1321 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     1381 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1441 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1501 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1561 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1621 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1681 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1741 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1801 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     1861 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     1921 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1981 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2041 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     2101 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     2161 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     2221 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2281 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2341 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2401 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2461 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2521 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2581 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     2641 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     2701 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     2761 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     2821 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     2881 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     2941 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3001 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     3061 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3121 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3181 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3241 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3301 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3541 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3601 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3661 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3721 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4201 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4621 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     5521 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     5581 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     5641 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     5701 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     5761 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5821 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5881 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     5941 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     6001 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     6061 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6121 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     6181 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     6241 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     6301 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     6361 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagc
//
