LOCUS       sequence-372936-        6938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6938)
  AUTHORS   Toth JI, Datta S, Athanikar JN, Freedman LP, Osborne TF
  TITLE     Selective coactivator interactions in gene activation by SREBP-1a 
            and -1c.
  JOURNAL   Mol Cell Biol. 2004 Sep . 24(18):8288-300.
  PUBMED    15340088
REFERENCE   2  (bases 1 to 6938)
  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 
            Biol. 2004 Sep . 24(18):8288-300."
COMMENT     Sequence Label: pcDNA3.1-2xFLAG-SREBP-1a
FEATURES             Location/Qualifiers
     source          1..6938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(61..80)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     307..327
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(336..764)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(446..467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     658..677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    810..1034
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1023..1040
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2611..2658)
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     primer_bind     complement(2687..2706)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2688..2706)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2751..2954)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2780..2800)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2955..3334
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3506..3525
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3600..3704
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3705..3773,3774..4565)
                     /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     complement(3923..3942)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4736..5324
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5225..5244
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5478..5495
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5612..5633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5648..5653,5654..5671,5672..5678)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5686..5702
                     /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)."
     primer_bind     5691..5713
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5710..5726
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5710..5726
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5775..5896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5786..5805)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5840..5859
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6070..6864)
                     /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(6181..6200)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6791..6810
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6931..6938,1..322))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gcagaggcgg
       61 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      301 ttccacaccc taactgacac acattccaca gaattaattc gcgttaaatt tttgttaaat
      361 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      421 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
      481 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
      541 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
      601 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
      661 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
      721 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga
      781 gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct
      841 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
      901 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
      961 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     1021 actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag agtcgacctg
     1081 caggcatgca agcttcaggg ccaggcggga gcggtccagc atgccccggc tgtgcagaga
     1141 cggccgctgc tctggctttg ccttgctgtc ctcaaagact gggctgtcag gctccgagtc
     1201 actgccactg ccaccgctgc cactgcccct gctgccaagg gacaaggggc tgctctggaa
     1261 aggtgagcca gcatccgagg ggggtggggt cagtgtgtcc tccacctcag tcttcacgcc
     1321 ctccatgagc acgtctgtgt tccctccact gccacaggcc gacaccagat ccttcagaga
     1381 tttgcttttg tggacagcag tgcgcagact taggttctcc tgcttgagtt tctggttgct
     1441 gtgttgcaga aagcgaatgt agtcgatggc cttgcgcaag acagcagatt tattcagctt
     1501 tgcctcagtg cccaccacca gatccttgag ctcaatgatt ttgtcattga tggaggagcg
     1561 gtagcgcttc tcaatggcgt tgtgggctgt gcgcttctct ccacggctct gggcagaggc
     1621 cggggccttg ctgccagctg cgagccggtt gataggcagc ttctccgcat ctacgaccag
     1681 tgggactgtt gccaagatgg ttccgccact caccagggtc ggcaaaggcc ctgtctgcac
     1741 agtggtgcca gagaccaggg gactgagacc tgccgccttc acagtggctc cgtccgtctt
     1801 catggctgtc agaagcagcg agtctgcctt gatgaagtgg ggctgcagca ggaccgggac
     1861 ctgctggatc tgcgaggtca cagtggtcgt tacaggggct gccgtggggg cagctgtgac
     1921 tgtcagtagc tgctggggga ccacactctg gacctgggtg tgcaaggaga cgggcgggac
     1981 ccctggtggg gaagccagtg gcaggccagg cagcggctgc tgggtgttcc cgggagggct
     2041 tcctgtagag aagcctcccg gagggctggg gtagcctaac acaggggtgg agctgaactg
     2101 cggtggggct ggggctggga agctctgtgg caggagggcc cctggcaggg gctgtggggt
     2161 gggggtctgc aggatgctca gtggcactga ctcttccttg ataccaggcc caggggagaa
     2221 agcgggcatg gacgggtaca tcttcaatgg agtgggtgca ggctggggag gggacagggg
     2281 tgagggcgct gcctgcggcc cgctcaggaa ggcttcaaga gaggagctca atgtggcagg
     2341 aggtggagac aagctgcctg gggagctggt atcggggctg gcagggtctg tgccccctgc
     2401 cccactccca gcatagggtg ggtcaaatag gccagggaag tcactgtctt ggttgttgat
     2461 aagctgaagc atgtcttcga tgtcggtcag cagcgccgcg tccagatcgc acggctcgcc
     2521 cagcgcctgc tccaaagccg cctcgctgaa gggtggctcg tccatggatc tgaattcgcc
     2581 cgggctggat ccgagctcgg taccaagctt cttatcgtcg tcatccttgt aatccttatc
     2641 gtcgtcatcc ttgtaatcca tagctcgagc tagccagctt gggtctccct atagtgagtc
     2701 gtattaattt cgataagcca gtaagcagtg ggttctctag ttagccagag agctctgctt
     2761 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     2821 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     2881 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     2941 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     3001 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     3061 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     3121 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     3181 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     3241 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     3301 gattactatt aataactagt caataatcaa tgtcaacgcg tatatctggc ccgtacatcg
     3361 cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc ggtcaagcct
     3421 tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca
     3481 gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3541 aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
     3601 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3661 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3721 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3781 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3841 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3901 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3961 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4021 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4081 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4141 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4201 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4261 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4321 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4381 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4441 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4501 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4561 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4621 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4681 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4741 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4801 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4861 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4921 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4981 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5041 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5101 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5161 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5221 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5281 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5341 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5401 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5461 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     5521 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5581 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     5641 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5701 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     5761 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     5821 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     5881 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     5941 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     6001 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     6061 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     6121 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     6181 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     6241 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     6301 atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc
     6361 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     6421 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     6481 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     6541 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     6601 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     6661 cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac
     6721 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     6781 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     6841 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatcc
     6901 gaaaatggat atacaagctc ccgggagctt tttgcaaa
//
