LOCUS       sequence-200857-        7270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of NR4A2 nuclear receptor subfamily 4 group A member 
            2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7270)
  AUTHORS   Shu Leong Ho
  TITLE     Shu Leong Ho Plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7270)
  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: 
            "unpublished"
COMMENT     Sequence Label: pcDNA3.1-hNurr1-NE
FEATURES             Location/Qualifiers
     source          1..7270
                     /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"
     primer_bind     complement(2627..2644)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2633..2857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2903..3331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2990..3009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3200..3221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3340..3360)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3345..3674
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3525..3660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3587..3606
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3741..4535
                     /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(3795..3814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4405..4424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4709..4830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4746..4765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4800..4819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4879..4895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4879..4895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4892..4914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4903..4919
                     /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(4927..4933,4934..4951,4952..4957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4972..4993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5110..5127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6040..6831,6832..6900))
                     /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     6663..6682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6901..7005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7080..7099)
                     /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 gctagcgttt aaacttaagc ttatgaccaa agaaaacccg cgtagcaacc aggaagaaag
      721 ctatgatgat aacgaaagcc cttgtgttca ggcgcagtat gggtcctcgc ctcaaggagc
      781 cagccccgct tctcagagct acagttacca ctcttcggga gaatacagct ccgatttctt
      841 aactccagag tttgtcaagt ttagcatgga cctcaccaac actgaaatca ctgccaccac
      901 ttctctcccc agcttcagta cctttatgga caactacagc acaggctacg acgtcaagcc
      961 accttgcttg taccaaatgc ccctgtccgg acagcagtcc tccattaagg tagaagacat
     1021 tcagatgcac aactaccagc aacacagcca cctgcccccc cagtctgagg agatgatgcc
     1081 gcactccggg tcggtttact acaagccctc ctcgcccccg acgcccacca ccccgggctt
     1141 ccaggtgcag cacagcccca tgtgggacga cccgggatct ctccacaact tccaccagaa
     1201 ctacgtggcc actacgcaca tgatcgagca gaggaaaacg ccagtctccc gcctctccct
     1261 cttctccttt aagcaatcgc cccctggcac cccggtgtct agttgccaga tgcgcttcga
     1321 cgggcccctg cacgtcccca tgaacccgga gcccgccggc agccaccacg tggtggacgg
     1381 gcagaccttc gctgtgccca accccattcg caagcccgcg tccatgggct tcccgggcct
     1441 gcagatcggc cacgcgtctc agctgctcga cacgcaggtg ccctcaccgc cgtcgcgggg
     1501 ctccccctcc aacgaggggc tgtgcgctgt gtgtggggac aacgcggcct gccaacacta
     1561 cggcgtgcgc acctgtgagg gctgcaaagg cttctttaag cgcacagtgc aaaaaaatgc
     1621 aaaatacgtg tgtttagcaa ataaaaactg cccagtggac aagcgtcgcc ggaatcgctg
     1681 tcagtactgc cgatttcaga agtgcctggc tgttgggatg gtcaaagaag tggttcgcac
     1741 agacagttta aaaggccgga gaggtcgttt gccctcgaaa ccgaagagcc cacaggagcc
     1801 ctctccccct tcgcccccgg tgagtctgat cagtgccctc gtcagggccc atgtcgactc
     1861 caacccggct atgaccagcc tggactattc caggttccag gcgaaccctg actatcaaat
     1921 gagtggagat gacacccagc atatccagca attctatgat ctcctgactg gctccatgga
     1981 gatcatccgg ggctgggcag agaagatccc tggcttcgca gacctgccca aagccgacca
     2041 agacctgctt tttgaatcag ctttcttaga actgtttgtc cttcgattag catacaggtc
     2101 caacccagtg gagggtaaac tcatcttttg caatggggtg gtcttgcaca ggttgcaatg
     2161 cgttcgtggc tttggggaat ggattgattc cattgttgaa ttctcctcca acttgcagaa
     2221 tatgaacatc gacatttctg ccttctcctg cattgctgcc ctggctatgg tcacagagag
     2281 acacgggctc aaggaaccca agagagtgga agaactgcaa aacaagattg taaattgtct
     2341 caaagaccac gtgactttca acaatggggg gttgaaccgc cccaattatt tgtccaaact
     2401 gttggggaag ctcccagaac ttcgtaccct ttgcacacag gggctacagc gcattttcta
     2461 cctgaaattg gaagacttgg tgccaccgcc agcaataatt gacaaacttt tcctggacac
     2521 tttacctttc taaggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag
     2581 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2641 tctagttgcc agccatctat tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2701 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2761 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2821 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2881 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3301 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3421 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3481 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3541 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3601 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3661 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3721 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3781 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3841 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3901 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3961 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4021 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4081 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4141 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4201 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4261 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4321 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4381 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4441 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4501 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4561 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4681 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4741 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4801 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4861 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7021 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7081 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7141 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7201 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     7261 tatacgcgtt
//
