LOCUS       sequence-338744-        4357 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 4357)
  TITLE     pCMV-STAT6-IRES-Neo
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4357)
  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            "Unpublished"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: p4xSTAT6-Luc2P
FEATURES             Location/Qualifiers
     source          1..4357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        206..237
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      264..273
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             270..1919
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             1923..2042
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2076..2124
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2280..2297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2451..3039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2531..2550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3239..4030,4031..4099))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4204..4252
                     /note="synthetic polyadenylation signal"
     misc_feature    4266..4357
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4306..4325
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccc gctgttgctc aatcgacttc ccaagaacag cgctgttgct
       61 caatcgactt cccaagaaca gcagggatct ggcctcggcg gccaagctcc gctgttgctc
      121 aatcgacttc ccaagaacag cgctgttgct caatcgactt cccaagaaca gcagggatct
      181 ggcctcggcg gccaagctta gacactagag ggtatataat ggaagctcga cttccagctt
      241 ggcaatccgg tactgttggt aaagccacca tggaagatgc caaaaacatt aagaagggcc
      301 cagcgccatt ctacccactc gaagacggga ccgccggcga gcagctgcac aaagccatga
      361 agcgctacgc cctggtgccc ggcaccatcg cctttaccga cgcacatatc gaggtggaca
      421 ttacctacgc cgagtacttc gagatgagcg ttcggctggc agaagctatg aagcgctatg
      481 ggctgaatac aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag ttcttcatgc
      541 ccgtgttggg tgccctgttc atcggtgtgg ctgtggcccc agctaacgac atctacaacg
      601 agcgcgagct gctgaacagc atgggcatca gccagcccac cgtcgtattc gtgagcaaga
      661 aagggctgca aaagatcctc aacgtgcaaa agaagctacc gatcatacaa aagatcatca
      721 tcatggatag caagaccgac taccagggct tccaaagcat gtacaccttc gtgacttccc
      781 atttgccacc cggcttcaac gagtacgact tcgtgcccga gagcttcgac cgggacaaaa
      841 ccatcgccct gatcatgaac agtagtggca gtaccggatt gcccaagggc gtagccctac
      901 cgcaccgcac cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc ggcaaccaga
      961 tcatccccga caccgctatc ctcagcgtgg tgccatttca ccacggcttc ggcatgttca
     1021 ccacgctggg ctacttgatc tgcggctttc gggtcgtgct catgtaccgc ttcgaggagg
     1081 agctattctt gcgcagcttg caagactata agattcaatc tgccctgctg gtgcccacac
     1141 tatttagctt cttcgctaag agcactctca tcgacaagta cgacctaagc aacttgcacg
     1201 agatcgccag cggcggggcg ccgctcagca aggaggtagg tgaggccgtg gccaaacgct
     1261 tccacctacc aggcatccgc cagggctacg gcctgacaga aacaaccagc gccattctga
     1321 tcacccccga aggggacgac aagcctggcg cagtaggcaa ggtggtgccc ttcttcgagg
     1381 ctaaggtggt ggacttggac accggtaaga cactgggtgt gaaccagcgc ggcgagctgt
     1441 gcgtccgtgg ccccatgatc atgagcggct acgttaacaa ccccgaggct acaaacgctc
     1501 tcatcgacaa ggacggctgg ctgcacagcg gcgacatcgc ctactgggac gaggacgagc
     1561 acttcttcat cgtggaccgg ctgaagagcc tgatcaaata caagggctac caggtagccc
     1621 cagccgaact ggagagcatc ctgctgcaac accccaacat cttcgacgcc ggggtcgccg
     1681 gcctgcccga cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg gaacacggta
     1741 aaaccatgac cgagaaggag atcgtggact atgtggccag ccaggttaca accgccaaga
     1801 agctgcgcgg tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc ggcaagttgg
     1861 acgcccgcaa gatccgcgag attctcatta aggccaagaa gggcggcaag atcgccgtga
     1921 attctcacgg cttccctccc gaggtggagg agcaggccgc cggcaccctg cccatgagct
     1981 gcgcccagga gagcggcatg gatagacacc ctgctgcttg cgccagcgcc aggatcaacg
     2041 tctaaggccg cgactctaga accggtcatg gccgcaataa aatatcttta ttttcattac
     2101 atctgtgtgt tggttttttg tgtgttcgaa ctagatgctg tcgaccgatg cccttgagag
     2161 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     2221 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
     2281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2881 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3181 agtatatatg agtaaacttg gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt
     3241 accagtgctt gatcagtgag gcaccgatct cagcgatctg cctatttcgt tcgtccatag
     3301 tggcctgact ccccgtcgtg tagatcacta cgattcgtga gggcttacca tcaggcccca
     3361 gcgcagcaat gatgccgcga gagccgcgtt caccggcccc cgatttgtca gcaatgaacc
     3421 agccagcagg gagggccgag cgaagaagtg gtcctgctac tttgtccgcc tccatccagt
     3481 ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt ttccgaagag
     3541 ttgtggccat tgctactggc atcgtggtat cacgctcgtc gttcggtatg gcttcgttca
     3601 actctggttc ccagcggtca agccgggtca catgatcacc catattatga agaaatgcag
     3661 tcagctcctt agggcctccg atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca
     3721 tggtaatggc agcactacac aattctctta ccgtcatgcc atccgtaaga tgcttttccg
     3781 tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg tatacggcga ccaagctgct
     3841 cttgcccggc gtctatacgg gacaacaccg cgccacatag cagtactttg aaagtgctca
     3901 tcatcgggaa tcgttcttcg gggcggaaag actcaaggat cttgccgcta ttgagatcca
     3961 gttcgatata gcccactctt gcacccagtt gatcttcagc atcttttact ttcaccagcg
     4021 tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac
     4081 gaaaatgttg gatgctcata ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt
     4141 actagtacgt ctctcaagga taagtaagta atattaaggt acgggaggta ttggacaggc
     4201 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     4261 gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
     4321 gtccccagtg caagtgcagg tgccagaaca tttctct
//
