LOCUS       sequence-275206-        5204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses CD4 with C-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5204)
  AUTHORS   Raissi AJ, Staudenmaier EK, David S, Hu L, Paradis S
  TITLE     Sema4D localizes to synapses and regulates GABAergic synapse 
            development as a membrane-bound molecule in the mammalian 
            hippocampus.
  JOURNAL   Mol Cell Neurosci. 2013 Nov;57:23-32. doi: 
            10.1016/j.mcn.2013.08.004. Epub 2013 Sep 10.
  PUBMED    24036351
REFERENCE   2  (bases 1 to 5204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Neurosci."; date: "2013-11"; volume: "57:23-32. doi"; pages: " 
            10.1016/j.mcn.2013.08.004. Epub 2013 Sep 10"
COMMENT     Sequence Label: pCMV-CD4-FLAG
FEATURES             Location/Qualifiers
     source          1..5204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      803..812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2279..2302
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2317..2451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2341..2360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2395..2414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2562..2578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2562..2578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2575..2597)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2586..2602
                     /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(join(2610..2616,2617..2634,2635..2640))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2655..2676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2793..2810)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2964..3552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3044..3063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3723..4514,4515..4583))
                     /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     4346..4365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4584..4688)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4756..4774
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4812..4834)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4934..4953
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5147..5169
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5161..5178
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5162..5178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcgggcccg ccgccaccat ggctgggatt ttctatttca tcctcttttc
      841 gtttctcttt ggaatttgcg acgctgtcac cggttctaga gggcccacca tggcatcaat
      901 gcagaagctg atctcagagg aggacctgct tatggccatg gaggcccgaa ttcggtcgac
      961 cgagatctta aagaaagtgg tgctgggcaa aaaaggggat acagtggaac tgacctgtac
     1021 agcttcccag aagaagagca tacaattcca ctggaaaaac tccaaccaga taaagattct
     1081 gggaattcag ggctccttct taactaaagg tccatccaag ctgaatgatc gcgctgactc
     1141 aagaagaagc ctttgggacc aaggaaactt ccccctgatc atcaagaatc ttaagataga
     1201 agactcagat acttacatct gtgaagtgga ggaccagaag gaggaggtgc aattgctagt
     1261 gttcggattg actgccaact ctgacaccca cctgcttcag gggcagagcc tgaccctgac
     1321 cttggagagc ccccctggta gtagcccctc agtgcaatgt aggagtccaa ggggtaaaaa
     1381 catacagggg gggaagaccc tctccgtgtc tcagctggag ctccaggata gtggcacctg
     1441 gacatgcact gtcttgcaga accagaagaa ggtggagttc aaaatagaca tcgtggtgct
     1501 agctttccag aaggcctcca gcatagtcta taagaaagag ggggaacagg tggagttctc
     1561 cttcccactc gcctttacag ttgaaaagct gacgggcagt ggcgagctgt ggtggcaggc
     1621 ggagagggct tcctcctcca agtcttggat cacctttgac ctgaagaaca aggaagtgtc
     1681 tgtaaaacgg gttacccagg accctaagct ccagatgggc aagaagctcc cgctccacct
     1741 caccctgccc caggccttgc ctcagtatgc tggctctgga aacctcaccc tggcccttga
     1801 agcgaaaaca ggaaagttgc atcaggaagt gaacctggtg gtgatgagag ccactcagct
     1861 ccagaaaaat ttgacctgtg aggtgtgggg acccacctcc cctaagctga tgctgagctt
     1921 gaaactggag aacaaggagg caaaggtctc gaagcgggag aaggcggtgt gggtgctgaa
     1981 ccctgaggcg gggatgtggc agtgtctgct gagtgactcg ggacaggtcc tgctggaatc
     2041 caacatcgag gttctgccca catggtccac cccggtgcag ccaatggccc tgattgtgct
     2101 ggggggcgtc gccggcctcc tgcttttcat tgggctaggc atcttcttct gtgtcaggtg
     2161 ccggcaccga aggcgccaag cagagcggat gtctcagatc aagagactcc tcagtgagaa
     2221 gaagacctgc cagtgccctc accggtttca gaagacatgt agccccattg gagcggccga
     2281 ctacaaggac gacgatgaca agtgagcggc cgcggggatc cagacatgat aagatacatt
     2341 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2401 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2461 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttcggatcct
     2521 ctagagtcga tctgcaggca tgctagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2581 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2641 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2701 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2761 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2821 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2881 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2941 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3001 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3061 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3121 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3181 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3241 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3301 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3361 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3421 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3481 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3541 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3601 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3661 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3721 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3781 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3841 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3901 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3961 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4021 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4081 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4141 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4201 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4261 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4321 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4381 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4441 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4501 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4561 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4621 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4681 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4741 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     4801 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     4861 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     4921 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     4981 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     5041 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     5101 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     5161 tgtaaaacga cggccagtga gttcgagctt gcatgcctgc aggt
//
