LOCUS       sequence-364650-        5610 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 5610)
  AUTHORS   Besser D, Bromberg JF, Darnell JE Jr, Hanafusa H
  TITLE     A single amino acid substitution in the v-Eyk intracellular domain 
            results in activation of Stat3 and enhances cellular transformation.
  JOURNAL   Mol Cell Biol 1999 Feb;19(2):1401-9.
  PUBMED    9891073
REFERENCE   2  (bases 1 to 5610)
  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"; date: "1999-02"; volume: "19(2)"; pages: "1401-9"
COMMENT     Sequence Label: 4xM67 pTATA TK-Luc
FEATURES             Location/Qualifiers
     source          1..5610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    10..144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        257..319
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             374..2026
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(440..458)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2089..2661
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2669..2688
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2764..2783)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2829..2884
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2883..2902)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3239..3255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3239..3255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3252..3274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3263..3279
                     /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(3287..3293,3294..3311,3312..3317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3332..3353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3470..3487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3641..4229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3721..3740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4400..5191,5192..5260))
                     /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     5023..5042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5261..5365)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5392..5526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 attgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
       61 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
      121 aatgtatctt atcatgtctg gatctcgaag cttgcatgcc tgcaggtcga cggttcccgt
      181 aaatgcatca ggttcccgta aatgcatcag gttcccgtaa atgcatcagg ttcccgtaaa
      241 tgatccagat gtccacttcg catattaagg tgacgcgtgt ggcctcgaac accgagcgac
      301 cctgcagcga cccgcttaac agcgtcaaca gcgtgccgca gatctcgaga tccattccgg
      361 tactgttggt aaaatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc
      421 tctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt
      481 tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt acgcggaata
      541 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca
      601 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt
      661 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa
      721 cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat
      781 tttgaacgtg caaaaaaaat taccaataat ccagaaaatt attatcatgg attctaaaac
      841 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt
      901 taatgaatac gattttgtac cagagtcctt tgatcgtgac aaaacaattg cactgataat
      961 gaattcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata gaactgcctg
     1021 cgtcagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc
     1081 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt
     1141 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttttacgatc
     1201 ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt cattcttcgc
     1261 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggggg
     1321 cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat
     1381 acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     1441 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     1501 ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca gaggacctat
     1561 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     1621 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatagttga
     1681 ccgcttgaag tctttaatta aatacaaagg atatcaggtg gcccccgctg aattggaatc
     1741 gatattgtta caacacccca acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga
     1801 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     1861 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     1921 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     1981 agagatcctc ataaaggcca agaagggcgg aaagtccaaa ttgtaaaatg taactgtatt
     2041 cagcgatgac gaaattctta gctattgtaa tatcctgaga acttcagggt gagtttgggg
     2101 acccttgatt gttctttctt tttcgctatt gtaaaattca tgttatatgg agggggcaaa
     2161 gttttcaggg tgttgtttag aatgggaaga tgtcccttgt atcaccatgg accctcatga
     2221 taattttgtt tctttcactt tctactctgt tgacaaccat tgtctcctct tattttcttt
     2281 tcattttctg taactttttc gttaaacttt agcttgcatt tgtaacgaat ttttaaattc
     2341 acttttgttt atttgtcaga ttgtaagtac tttctctaat cacttttttt tcaaggcaat
     2401 cagggtatat tatattgtac ttcagcacag ttttagagaa caattgttat aattaaatga
     2461 taaggtagaa tatttctgca tataaattct ggctggcgtg gaaatattct tattggtaga
     2521 aacaactaca tcctggtcat catcctgcct ttctctttat ggttacaatg atatacactg
     2581 tttgagatga ggataaaata ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat
     2641 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     2701 tttggcaaag aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     2761 ggatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     2821 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     2881 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     2941 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     3001 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     3061 gacttgaggt tagatttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     3121 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgacc actcccagtc
     3181 atagctatcc ctcttctctt atggagatcc cgggtaccga gctcgaattc gtaatcatgt
     3241 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3301 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3541 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4021 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4081 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4261 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4321 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4381 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4441 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4501 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4561 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4621 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4681 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4741 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4801 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4861 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4921 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4981 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5041 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5101 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5161 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5221 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5281 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5341 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaacttgttt
     5401 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5461 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5521 tggatcagcc atatcacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     5581 cccctgaacc tgaaacataa aatgaatgca
//
