LOCUS       Exported                6430 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Human CXCR4 coding sequence in vector for in vitro transcription and
            protein synthesis, with T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pET-24a-CXCR4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6430)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 6430)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6430
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             89..121
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             137..157
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1229..1249
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1250..1273
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1295..1312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1365..1383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1379..1426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1463..1918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1550..1569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1760..1781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2010..2825)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2733..2752
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2817..2836
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2947..3535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3436..3455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3689..3706
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3721..3860
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3786..3805)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3946..3968
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3962..4153)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4962..6044)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6006..6025
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6045..6122)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6328..6347
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggctagcatg actggtggac agcaaatggg
      121 tcgcggatcc gaattcgaaa acctgtattt tcagggcccg atggaaggaa tctctattta
      181 tacctctgat aactatactg aagaaatggg atccggagat tacgattcca tgaaagaacc
      241 ttgtttccgt gaggaaaacg ccaacttcaa caaaatattc cttccaacta tctattccat
      301 catttttttg accggaatcg ttggtaatgg actagttatt ttggttatgg gataccaaaa
      361 aaaacttcgt tccatgactg ataagtacag acttcatctt tctgttgctg atttgctttt
      421 cgttattacc cttcctttct gggccgtcga cgctgtcgcc aattggtact ttggaaattt
      481 cctttgtaag gccgttcacg ttatttatac cgtcaacctt tactcctcag tactgattct
      541 tgcttttatt tcccttgata gataccttgc catcgtgcat gccactaact ctcagagacc
      601 tagaaagctt cttgccgaga aagtcgtcta cgtaggagtt tggattccag ctcttttgtt
      661 gaccattcca gatttcattt ttgctaacgt ctccgaggct gatgatcgtt acatttgtga
      721 ccgtttttac ccaaatgacc tttgggttgt cgttttccaa tttcaacata tcatggtcgg
      781 acttattttg ccaggaattg tcattctttc ctgttactgc atcatcatct ccaagctttc
      841 tcattctaaa ggtcaccaga aaagaaaagc cttgaagacc accgtcatct tgattcttgc
      901 cttttttgct tgttggcttc catattatat cggtatttcc atcgattcct tcatccttct
      961 tgaaattatt aaacaaggat gtgaatttga gaacactgtc cataagtgga tctccattac
     1021 cgaggccttg gctttcttcc attgctgtct taaccctatt ttgtatgcct ttttgggcgc
     1081 caaattcaag acttccgctc aacacgcctt gacctctgtt tctcgtggat cttctcttaa
     1141 gattttgtct aaaggaaaac gtggaggaca ctcctctgtc tccaccgaat ctgaatcctc
     1201 ttcctttcat tcctctggat tgggcccaga aaacctgtac ttccagggtc accatcatca
     1261 ccaccatcat cactaataag cggccgcact cgagcaccac caccaccacc actgagatcc
     1321 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1381 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1441 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1501 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1561 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1621 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1681 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1741 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1801 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1861 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1921 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1981 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     2041 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     2101 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     2161 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     2221 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     2281 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     2341 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     2401 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     2461 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     2521 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     2581 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     2641 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     2701 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     2761 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     2821 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     2881 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3061 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3301 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3361 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3541 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3601 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3661 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3721 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3781 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3841 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3901 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3961 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4021 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4081 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4141 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     4201 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     4261 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     4321 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     4381 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     4441 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     4501 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4561 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4621 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     4681 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     4741 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     4801 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     4861 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     4921 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4981 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5041 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     5101 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     5161 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     5221 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     5281 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     5341 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     5401 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     5461 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     5521 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5581 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5641 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5701 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5761 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5821 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5881 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5941 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     6001 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     6061 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     6121 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     6181 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     6241 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     6301 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     6361 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     6421 cccgcgaaat
//
