LOCUS       sequence-150366-        8149 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 8149)
  AUTHORS   Zhong Z, Wen Z, Darnell JE Jr
  TITLE     Stat3 and Stat4: members of the family of signal transducers and 
            activators of transcription.
  JOURNAL   Proc Natl Acad Sci U S A 1994 May 24;91(11):4806-10.
  PUBMED    7545930
REFERENCE   2  (bases 1 to 8149)
  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: "Proc Natl 
            Acad Sci U S A"; date: "1994-05-24"; volume: "91(11)"; pages: 
            "4806-10"
COMMENT     Sequence Label: Stat4 pRc/CMV
FEATURES             Location/Qualifiers
     source          1..8149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        117..496
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        497..700
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     651..671
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     745..764
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        745..763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     854..870
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     854..870
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3459..3475)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3460..3476)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3490..3508)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3491..3508)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3528..3545)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3534..3758
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3804..4317
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3891..3910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4101..4122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4340..4360)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4345..4674
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4525..4660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4741..5535
                     /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(4795..4814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5405..5424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5709..5843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5746..5765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5800..5819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5874..5890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5874..5890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5887..5909)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5898..5914
                     /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(5922..5928,5929..5946,5947..5952))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5967..5988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6105..6122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6276..6864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6356..6375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7035..7826,7827..7895))
                     /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     7658..7677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7896..8000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8075..8094)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
       61 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      121 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      181 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      241 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      481 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      541 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      601 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      661 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      721 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
      781 ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattctgcag atatccatca
      841 cactggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcc tgccactacc
      901 tggacggaga gagagagagc cagcatgtct cagtggaatc aagtccaaca attagaaatc
      961 aagtttttgg agcaagtaga tcagttctat gatgacaact ttcctatgga aatccggcat
     1021 ctgctagctc agtggattga gactcaagac tgggaagtag cttctaacaa tgaaactatg
     1081 gcaacaattc tgcttcaaaa cttactaata caattggatg aacagttggg gcgggtttcc
     1141 aaagaaaaaa atctgctatt gattcacaat ctaaagagaa ttagaaaagt tcttcagggc
     1201 aagtttcatg gaaatccaat gcatgtagct gtggtaattt caaattgctt aagggaagag
     1261 aggagaatat tggctgcagc caacatgcct atccagggac ctctggagaa atccttacag
     1321 agttcttcag tttctgaaag acaaaggaat gtggaacaca aagtgtctgc cattaaaaac
     1381 agtgtgcaga tgacagaaca agataccaaa tacttagaag acctgcaaga tgagtttgac
     1441 tacaggtata aaacaattca gacaatggat cagggtgaca aaaacagtat cctggtgaac
     1501 caggaagttt tgacgctgca agaaatgctt aatagtctgg acttcaagag aaaggaagca
     1561 ctcagtaaga tgacgcagat agtgaacgag acagacctgc tcatgaacag catgcttcta
     1621 gaagagctgc aggactggaa aaagcggcag cagattgcct gcattggtgg cccgctccac
     1681 aatgggctgg accagcttca gaactgcttt accctactgg cagagagtct tttccaactc
     1741 agacagcaac tggagaaact acaggagcaa tctactaaaa tgacctatga aggggatccc
     1801 atccctgctc aaagagcaca cctcctggaa agagctacct tcctgatcta caaccttttc
     1861 aagaactcat ttgtggtcga gcgacagcca tgcatgccaa cgcaccctca gaggccgatg
     1921 gtacttaaaa ccctcattca gttcactgta aaactgagat tactaataaa attgccggaa
     1981 ctaaactatc aggtgaaagt aaaggcgtcc attgacaaga atgtttcaac tctaagcaat
     2041 agaagatttg tgctttgtgg aactcacgtc aaagctatgt ccagtgagga atcttccaat
     2101 gggagcctct cagtggagtt tagacatttg caacccaagg agatgaagtg cagtactggg
     2161 agtaaaggaa acgagggctg ccacatggtg acagaggagt tgcattccat aacctttgag
     2221 acccagatct gcctctatgg cctcaccatt aacctagaga ccagctcatt acctgtcgtg
     2281 atgatttcta atgtcagcca actacctaat gcatgggcat ccatcatttg gtacaatgta
     2341 tcaactaacg actcccagaa cttggttttc tttaataacc ctccatctgt cactttgggc
     2401 caactcctgg aagtgatgag ctggcaattt tcatcctatg tcggtcgtgg ccttaattca
     2461 gagcagctca acatgctggc agagaagctc acagttcagt ctaactacaa tgatggtcac
     2521 ctcacctggg ccaagttctg caaggaacat ttgcctggca aaacatttac cttctggact
     2581 tggcttgaag caatattgga cctaattaaa aaacatattc ttcccctctg gattgatggg
     2641 tacatcatgg gatttgttag taaagagaag gaacggcttc tgctcaaaga taaaatgcct
     2701 gggacatttt tgttaagatt cagtgagagc catcttggag ggataacctt cacctgggtg
     2761 gaccaatctg aaaatggaga agtgagattc cactctgtag aaccctacaa caaagggaga
     2821 ctgtcggctc tgccgttcgc tgacatcctg cgagactaca aggttatcat ggctgaaaac
     2881 atccctgaaa accctctgaa gtacctctac cctgacattc ccaaagacaa agcctttggc
     2941 aaacactaca gctcccagcc gtgcgaagtc tcaagaccaa ccgaacgggg agacaagggt
     3001 tacgtcccct ctgtttttat ccccatttca acaatccgaa gcgattccac ggagccacaa
     3061 tctccttcag accttctccc catgtctcca agtgcatatg ctgtgctgag agaaaacctg
     3121 agcccaacga caattgaaac tgcaatgaat tccccatatt ctgctgaatg acggtgcaaa
     3181 cggacacttt aaagaaggaa gcagatgaaa ctggagagtg ttctttacca tagatcacaa
     3241 tttatttctt cggctttgta aataccagtt tctagaaaat ggtaggaagt tggaagctct
     3301 cttctcactt ggtgccactc ccagcaagga gcgctgtgac taaaatgcta aaaaacctgc
     3361 aagaaaagac agctttaaga aaccagtgtt ggtaacaata taacagagga ctcctttgaa
     3421 aaaaaaaaaa aaaaaaagga attcgatatc aagcttatcg ataccgtcga cctcgagggg
     3481 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     3541 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3601 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3661 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3721 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3781 ctggggctcg aggggggatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3841 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3901 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3961 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4021 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4081 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4141 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4201 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     4261 aatatttgct tatacaatct tcctgttttt ggggcttttc tgattatcaa ccggggtggg
     4321 taccgagctc gaattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4381 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4441 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4501 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4561 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4621 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4681 ccgggagctt ggatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4741 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4801 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4861 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4921 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4981 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5041 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5101 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5161 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5221 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5281 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5341 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5401 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5461 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5521 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5581 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5641 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5701 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5761 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5821 atgtatctta tcatgtctgg atccgtcgac ctcgagagct tggcgtaatc atggtcatag
     5881 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5941 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6001 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6061 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     8041 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     8101 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtg
//
