LOCUS       sequence-381367-        6679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nuclear localization signal (NLS), C-terminal myc-(His)6 
            fusion of human gene or portion of gene in human cell culture lines.
            pcDNA4.1-based vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6679)
  TITLE     Davis Lab plasmids
REFERENCE   2  (bases 1 to 6679)
  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"
COMMENT     Sequence Label: PRPF4.001.522.Y32A.pc4
FEATURES             Location/Qualifiers
     source          1..6679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2368..2397
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2413..2430
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2453..2470)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2459..2683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2729..3157
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2816..2835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3026..3047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3166..3186)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3171..3500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3351..3486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3413..3432
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3548..3595
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3614..3988
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4118..4239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4155..4174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4209..4228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4288..4304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4288..4304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4301..4323)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4312..4328
                     /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(4336..4342,4343..4360,4361..4366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4381..4402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4519..4536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4690..5278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4770..4789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5449..6240,6241..6309))
                     /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     6072..6091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6310..6414)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6489..6508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtccat gggaaaaaaa acaggaaaga acagaaaatt gaagtccaag
      721 cgggtcaaac ctcgatggaa ttctgcagat atccagcaca gtggcggccg catggcttcc
      781 tcgcgagcct cttccacgca ggcaaccaaa actaaagcac ccgacgactt agttgctccg
      841 gtcgtgaaga aaccacacat ctatgctgga agtttggaag agaaggagag ggagcgtctg
      901 gccaaaggag agtctgggat tttggggaaa gacggactta aagcagggat cgaagctgga
      961 aatattaata taacctctgg agaagtgttt gaaattgaag agcatatcag cgagcgacag
     1021 gcagaagtat tggctgagtt tgagagaagg aagcgagccc ggcagatcaa tgtttccaca
     1081 gatgactcag aggtcaaagc ttgccttaga gccttggggg aacccatcac actttttgga
     1141 gagggtcctg ctgaaagaag agaaaggtta agaaatatcc tctcagttgt cggtactgat
     1201 gccttgaaaa agaccaaaaa ggatgatgag aagtctaaaa agtccaaaga agagtatcag
     1261 caaacctggt atcatgaagg accaaatagc ttgaaggtgg caagactatg gattgctaat
     1321 tattcgttgc ccagggcaat gaaacgcttg gaagaggccc gactccataa ggagattcct
     1381 gagacaacaa ggacctccca gatgcaagag ctgcacaagt ctctccggtc tttgaataat
     1441 ttttgcagtc agattgggga tgatcggcct atctcctact gtcactttag tcccaattcc
     1501 aagatgctgg ccacagcttg ttggagtggg ctttgcaagc tctggtctgt tcctgattgc
     1561 aacctccttc acactcttcg agggcataac acaaatgtag gagcaattgt attccatccc
     1621 aaatccactg tctccttgga cccaaaagat gtcaacctgg cctcttgtgc ggctgatggc
     1681 tctgtgaagc tttggagtct cgacagtgat gaaccagtgg cagatattga aggccataca
     1741 gtgcgtgtgg cgcgggtaat gtggcatcct tcaggacgtt tcctgggcac cacctgctat
     1801 gaccgttcat ggcgcttatg ggatttggag gctcaagagg agatcctgca tcaggaaggc
     1861 catagcatgg gtgtgtatga cattgccttc catcaagatg gctctttggc tggcactggg
     1921 ggactggatg catttggtcg agtttgggac ctacgcacag gacgttgtat catgttctta
     1981 gaaggccacc tgaaagaaat ctatggaata aatttctccc ccaatggcta tcacattgca
     2041 accggcagtg gtgacaacac ctgcaaagtg tgggacctcc gacagcggcg ttgcgtctac
     2101 accatccctg ctcatcagaa cttagtgact ggtgtcaagt ttgagcctat ccatgggaac
     2161 ttcttgctta ctggtgccta tgataacaca gccaagatct ggacgcaccc aggctggtcc
     2221 ccgctgaaga ctctggctgg ccacgaaggc aaagtgatgg gcctagatat ttcttccgat
     2281 gggcagctca tagccacttg ctcatatgac aggaccttca agctgtggat ggctgaaggc
     2341 cgctcgagtc tagagggccc gcggttcgaa caaaaactca tctcagaaga ggatctgaat
     2401 atgcataccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     2461 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2521 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2581 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2641 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2701 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2881 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2941 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3061 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3121 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3181 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3241 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3301 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3361 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3421 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3481 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3541 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3601 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     3661 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     3721 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     3781 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     3841 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     3901 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     3961 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     4021 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4081 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4141 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4201 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4261 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4441 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5101 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6421 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     6481 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     6541 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     6601 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     6661 cgggccagat atacgcgtt
//
