LOCUS       sequence-266317-        7526 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 7526)
  AUTHORS   Macian F, Garcia-Rodriguez C, Rao A
  TITLE     Gene expression elicited by NFAT in the presence or absence of 
            cooperative recruitment of Fos and Jun.
  JOURNAL   EMBO J. 2000 Sep 1. 19(17):4783-95.
  PUBMED    10970869
REFERENCE   2  (bases 1 to 7526)
  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: "EMBO J. 
            2000 Sep 1. 19(17):4783-95."
COMMENT     Sequence Label: IL-4 promoter luciferase
FEATURES             Location/Qualifiers
     source          1..7526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             1013..2665
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1079..1097)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2796..2861
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2991..3011
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3436..3570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4130..4150)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4130..4148)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4169..4185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4169..4185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4182..4204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4193..4209
                     /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(4217..4223,4224..4241,4242..4247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4262..4283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4400..4417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4571..5159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4651..4670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5330..6121,6122..6190))
                     /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     5953..5972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6191..6295)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6322..6778
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6409..6428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6619..6640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6904..6926
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6918..6935
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6919..6935
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6945..6964
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6945..6963
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6986..7005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    7299..7433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
       61 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      121 tatcatgtct ggatccaagc tcagatccaa gcttgtcgac ccgggtaccg agctcgagat
      181 ccacactgat gctgtagtgc acatagatac acacatgctc acatgaagta atttaaaaaa
      241 attttttttt aaatcagcca tttctcaggc ttctgtctaa ggtaggaaaa atcttcaacc
      301 tagcccagaa cctccatata ggtaaagcct cattccatgg tcctgcctgc cccactccat
      361 gtcacctctc tgtctccaaa gaccacaaac ttgtaagatc agctggtcta ggatgcgaga
      421 aggtctgcct ccatcatcct tctatgaggt aagaccccag agtcagcttt cccaagatat
      481 cagagtttcc aaggggcccc catagcagga agcagctagg cccaggtgtg ctcaaggcag
      541 actttcttga tattactctg tctttcccca gggcgacacc agcaccctcg gacacctgtg
      601 acctcttcct tctctgcagg aggagagcca gtggcaaccc tacgctgata agattagtct
      661 gaaaggccga ttatggtgta atttcctatg ctgaaacttt gtagatttaa aaaaaagggg
      721 ggggaggggt gtttcatttt ccaattggtc tgatttcaca ggaaaattta cctgtttctc
      781 ttttttctcc tggaagagag gtgctgattg gcccagaata actgacaatc tggtgtaata
      841 aaattttcca atgtaaactc attttccctt ggtttcagca actttaactc tatatataga
      901 gagacctctg ccagcattgc attgttagca tctcttgata aacttaattg tctctcgtca
      961 ctgacggcac agagctagga tctgagcttg gcattccggt actgttggta aaatggaaga
     1021 cgccaaaaac ataaagaaag gcccggcgcc attctatcct ctagaggatg gaaccgctgg
     1081 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
     1141 agatgcacat atcgaggtga acatcacgta cgcggaatac ttcgaaatgt ccgttcggtt
     1201 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
     1261 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
     1321 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgaaca tttcgcagcc
     1381 taccgtagtg tttgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaaatt
     1441 accaataatc cagaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
     1501 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtacc
     1561 agagtccttt gatcgtgaca aaacaattgc actgataatg aattcctctg gatctactgg
     1621 gttacctaag ggtgtggccc ttccgcatag aactgcctgc gtcagattct cgcatgccag
     1681 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
     1741 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
     1801 cttaatgtat agatttgaag aagagctgtt tttacgatcc cttcaggatt acaaaattca
     1861 aagtgcgttg ctagtaccaa ccctattttc attcttcgcc aaaagcactc tgattgacaa
     1921 atacgattta tctaatttac acgaaattgc ttctgggggc gcacctcttt cgaaagaagt
     1981 cggggaagcg gttgcaaaac gcttccatct tccagggata cgacaaggat atgggctcac
     2041 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     2101 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     2161 cgttaatcag agaggcgaat tatgtgtcag aggacctatg attatgtccg gttatgtaaa
     2221 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     2281 agcttactgg gacgaagacg aacacttctt catagttgac cgcttgaagt ctttaattaa
     2341 atacaaagga tatcaggtgg cccccgctga attggaatcg atattgttac aacaccccaa
     2401 catcttcgac gcgggcgtgg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     2461 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     2521 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     2581 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     2641 gaagggcgga aagtccaaat tgtaaaatgt aactgtattc agcgatgacg aaattcttag
     2701 ctattgtaat actctagagg atctttgtga aggaacctta cttctgtggt gtgacataat
     2761 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat
     2821 aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg
     2881 atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa
     2941 tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga
     3001 agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg
     3061 ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg
     3121 cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt aggcataaca
     3181 gttataatca taacatactg ttttttctta ctccacacag gcatagagtg tctgctatta
     3241 ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg gttaataagg
     3301 aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac atttgtagag
     3361 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3421 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3481 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3541 ctcatcaatg tatcttatca tgtctggatc gatccccagg aagctcctct gtgtcctcat
     3601 aaaccctaac ctcctctact tgagaggaca ttccaatcat aggctgccca tccaccctct
     3661 gtgtcctcct gttaattagg tcacttaaca aaaaggaaat tgggtagggg tttttcacag
     3721 accgctttct aagggtaatt ttaaaatatc tgggaagtcc cttccactgc tgtgttccag
     3781 aagtgttggt aaacagccca caaatgtcaa cagcagaaac atacaagctg tcagctttgc
     3841 acaagggccc aacaccctgc tcatcaagaa gcactgtggt tgctgtgtta gtaatgtgca
     3901 aaacaggagg cacattttcc ccacctgtgt aggttccaaa atatctagtg ttttcatttt
     3961 tacttggatc aggaacccag cactccactg gataagcatt atccttatcc aaaacagcct
     4021 tgtggtcagt gttcatctgc tgactgtcaa ctgtagcatt ttttggggtt acagtttgag
     4081 caggatattt ggtcctgtag tttgctaaca caccctccag cttttgttcc ctttagtgag
     4141 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6301 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6361 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6541 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6601 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6661 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaaat
     6721 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     6781 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6841 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6901 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     6961 agggcgaatt ggggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7021 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     7081 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     7141 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     7201 ggatcagcca tatcacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     7261 ccctgaacct gaaacataaa atgaatgcaa ttgttgttaa cttgtttatt gcagcttata
     7321 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7381 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcagccata
     7441 tcacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     7501 aacataaaat gaatgcaatt gttgtt
//
