LOCUS       sequence-173345-        6674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of the transcription factor Brn3a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  AUTHORS   Schmidt K, Joyce CE, Buquicchio F, Brown A, Ritz J, Distel RJ, Yoon 
            CH, Novina CD
  TITLE     The lncRNA SLNCR1 Mediates Melanoma Invasion through a Conserved 
            SRA1-like Region.
  JOURNAL   Cell Rep. 2016 May 31;15(9):2025-37. doi: 
            10.1016/j.celrep.2016.04.018. Epub 2016 May 19.
  PUBMED    27210747
REFERENCE   2  (bases 1 to 6674)
  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"; doi: 
            "10.1016/j.celrep.2016.04.018"; journalName: "Cell Rep"; date: 
            "2016-05-31- 31"; volume: "15"; issue: "9"; pages: "2025-37"
COMMENT     Sequence Label: pCDNA3.1 (-) + codon optimized Brn3a
FEATURES             Location/Qualifiers
     source          1..6674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             643..669
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(1651..1668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1657..1881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1927..2355
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2014..2033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2224..2245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2364..2384)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2369..2698
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2549..2684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2611..2630
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2765..3559
                     /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(2819..2838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3429..3448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3733..3854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3770..3789)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3824..3843
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3903..3919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3903..3919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3916..3938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3927..3943
                     /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(3951..3957,3958..3975,3976..3981))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3996..4017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4134..4151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4305..4893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4385..4404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5064..5855,5856..5924))
                     /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     5687..5706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5925..6029)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6104..6123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6295..6674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacgggccc
      301 tctagactcg agcggccgcc actgtgctgg atatctgcag aattcatgat gtctatgaac
      361 tctaagcagc ctcactttgc aatgcaccca acactgccag agcacaagta ccctagcctg
      421 cactcatctt ctgaggcaat tagacgcgct tgtctcccta ctcctcccct ccagtcaaat
      481 ctgttcgcat cactcgacga gacactcctg gcaagggcag aggccctcgc cgctgtggac
      541 attgccgtgt cccagggaaa gtctcacccg tttaagcccg acgctacata ccacactatg
      601 aactcagtgc catgtacatc aacttcaacc gtgcctctcg cacaccacca ccaccaccac
      661 caccaccacc aggctttaga acccggcgat ctcctggatc acatttctag cccatctctg
      721 gctctgatgg ccggggcagg tggagccgga gcagctgcgg gcggtggcgg agcacacgat
      781 ggacccggcg gcggcggtgg acctggcggt ggcggcggac ctggaggcgg ccccggtgga
      841 ggcggaggag gtgggcctgg cggcggaggc ggcgggcccg gcggaggact cctgggaggt
      901 tctgcacacc ctcaccccca catgcactct ctgggacacc tgtcccaccc agcagcagct
      961 gctgctatga atatgccttc aggactgcct caccccggac tcgtcgccgc cgcagcacac
     1021 cacggggcag ctgctgctgc cgccgccgcg gctgccggac aggtggccgc cgcaagcgcc
     1081 gcagctgccg tcgtcggcgc cgctggactg gcttctattt gtgattccga taccgatcca
     1141 cgcgaattgg aggctttcgc tgagcggttt aagcagagac ggattaagct cggagtgaca
     1201 caggccgacg tcgggtccgc actggctaac ctcaagattc ccggcgtcgg gtcactgtca
     1261 cagtctacta tttgtaggtt tgagtcactg actctgtctc acaacaatat gattgccctc
     1321 aagccaatcc tccaggcatg gctcgaagaa gctgagggcg ctcagcgcga gaagatgaat
     1381 aagccagaac tctttaatgg gggcgagaag aagagaaaga gaacttccat tgctgctcct
     1441 gagaagcggt ccttagaggc ctacttcgca gtccagccaa gaccttcatc cgagaagatt
     1501 gctgctattg ctgagaagct cgatctcaag aagaacgtgg tgcgcgtgtg gttttgtaat
     1561 cagagacaga agcagaagcg catgaagttt agcgctacat actaactcga gtaagtatcc
     1621 cgggaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat
     1681 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1741 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     1801 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     1861 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     1921 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1981 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2101 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2221 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2281 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2341 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2401 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2461 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2521 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2581 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     2641 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2701 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     2761 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2821 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2881 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2941 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3001 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3061 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3121 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3181 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3241 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3301 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3361 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3421 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3481 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3541 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3601 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3661 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3721 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3781 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3841 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     3901 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3961 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4021 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4081 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4141 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4201 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4261 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4321 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4381 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4441 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4501 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4561 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4621 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4681 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4741 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4801 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4861 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4921 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4981 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5041 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5101 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5161 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5221 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5281 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5341 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5401 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5461 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5521 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5581 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5641 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5701 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5761 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5821 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5881 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5941 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6001 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6061 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6121 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6181 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     6241 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     6301 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     6361 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6421 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6481 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6541 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6601 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6661 tcgctattac catg
//
