LOCUS       sequence-254628-        6494 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of HuR (wild type).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6494)
  TITLE     Dias lab diverse plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6494)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1D V5-His-TOPO-HuR.wt
FEATURES             Location/Qualifiers
     source          1..6494
                     /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             1758..1799
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1809..1826
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1849..1866)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1855..2079
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2125..2553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2212..2231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2422..2443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2562..2582)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2567..2896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2747..2882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2809..2828
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2963..3757
                     /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(3017..3036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3627..3646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3933..4054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3970..3989)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4024..4043
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4103..4119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4103..4119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4116..4138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4127..4143
                     /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(4151..4157,4158..4175,4176..4181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4196..4217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4334..4351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4505..5093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4585..4604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5264..6055,6056..6124))
                     /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     5887..5906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6125..6229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6304..6323)
                     /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 cttggtaccg agctcggatc cagtaccctt caccatgtct aatggttatg
      721 aagaccacat ggccgaagac tgcaggggtg acatcgggag aacgaatttg atcgtcaact
      781 acctccctca gaacatgacc caggatgagt tacgaagcct gttcagcagc attggtgaag
      841 ttgaatctgc aaaacttatt cgggataaag tagcaggaca cagcttgggc tatggctttg
      901 tgaactacgt gaccgcgaag gatgcagaga gagcgatcaa cacgctgaac ggcttgaggc
      961 tccagtcaaa aaccattaag gtgtcgtatg ctcgcccgag ctcagaggtg atcaaagacg
     1021 ccaacttgta catcagcggg ctcccgcgga ccatgaccca gaaggacgta gaagacatgt
     1081 tctctcggtt tgggcggatc atcaactcgc gggtcctcgt ggatcagact acaggtttgt
     1141 ccagaggggt tgcgtttatc cggtttgaca aacggtcgga ggcagaagag gcaattacca
     1201 gtttcaatgg tcataaaccc ccaggttcct ctgagcccat cacagtgaag tttgcagcca
     1261 accccaacca gaacaaaaac gtggcactcc tctcgcagct gtaccactcg ccagcgcgac
     1321 ggttcggagg ccccgttcac caccaggcgc agagattcag gttctccccc atgggcgtcg
     1381 atcacatgag cgggctctct ggcgtcaacg tgccaggaaa cgcctcctcc ggctggtgca
     1441 ttttcatcta caacctgggg caggatgccg acgaggggat cctctggcag atgtttgggc
     1501 cgtttggtgc cgtcaccaat gtgaaagtga tccgcgactt caacaccaac aagtgcaaag
     1561 ggtttggctt tgtgaccatg acaaactatg aagaagccgc gatggccata gccagcctga
     1621 acggctaccg cctgggggac aaaatcttac aggtttcctt caaaaccaac aagtcccaca
     1681 aatttaaggg tcaagacaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     1741 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     1801 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     1861 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1921 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1981 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2041 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2101 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2161 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2221 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2281 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2341 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2401 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2461 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2521 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     2581 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2641 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2701 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2761 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2821 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2881 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     2941 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3001 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3061 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3121 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3181 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3241 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3301 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3361 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3421 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3481 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3541 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3601 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3661 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3721 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     3781 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3841 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3901 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3961 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4021 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4081 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4141 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4201 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4261 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4321 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4381 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4441 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4501 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4561 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4621 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4681 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4741 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4801 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4861 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4921 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4981 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5041 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5101 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5161 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5221 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5281 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5341 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5401 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5461 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5521 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5581 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5641 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5701 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5761 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5821 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5881 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5941 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6001 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6061 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6121 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6181 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6241 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     6301 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     6361 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     6421 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     6481 cagatatacg cgtt
//
