LOCUS       sequence-381459-        4990 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 4990)
  AUTHORS   Ueki K, Kondo T, Kahn CR
  TITLE     Suppressor of cytokine signaling 1 (SOCS-1) and SOCS-3 cause insulin
            resistance through inhibition of tyrosine phosphorylation of insulin
            receptor substrate proteins by discrete mechanisms.
  JOURNAL   Mol Cell Biol. 2004 Jun . 24(12):5434-46.
  PUBMED    15169905
REFERENCE   2  (bases 1 to 4990)
  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 Jun . 24(12):5434-46."
COMMENT     Sequence Label: pCMV SOCS-3
FEATURES             Location/Qualifiers
     source          1..4990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        616..634
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             678..701
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1415..1431)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1416..1432)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1487..1506)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1488..1506)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1780..1901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1817..1836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1871..1890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1908..2363)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2045..2066)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2257..2276
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2390..2494
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2496..2853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2496..2687
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2519..2539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2704..2839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2888..3682
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2888..3682
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2942..2961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3552..3571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3870..3889)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3914..3961
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4290..4878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4779..4798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcga
      601 ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggagc tccaccgcgg
      661 tggcggccgc caccatggat tacaaggatg acgacgataa gagcccgggc ggatccatgg
      721 tcacccacag caagtttccc gccgccggga tgagccgccc cctggacacc agcctgcgcc
      781 tcaagacctt cagctccaaa agcgagtacc agctggtggt gaacgccgtg cgcaagctgc
      841 aggagagcgg attctactgg agcgccgtga ccggcggcga ggcgaacctg ctgctcagcg
      901 ccgagcccgc gggcaccttt cttatccgcg acagctcgga ccagcgccac ttcttcacgt
      961 tgagcgtcaa gacccagtcg gggaccaaga acctacgcat ccagtgtgag gggggcagct
     1021 tttcgctgca gagtgacccc cgaagcacgc agccagttcc ccgcttcgac tgtgtactca
     1081 agctggtgca ccactacatg ccgcctccag ggaccccctc cttttctttg ccacccacgg
     1141 aaccctcgtc cgaagttccg gagcagccac ctgcccaggc actccccggg agtaccccca
     1201 agagagctta ctacatctat tctgggggcg agaagattcc gctggtactg agccgacctc
     1261 tctcctccaa cgtggccacc ctccagcatc tttgtcggaa gactgtcaac ggccacctgg
     1321 actcctatga gaaagtgacc cagctgcctg gacccattcg ggagttcctg gatcagtatg
     1381 atgctccact ttaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc
     1441 ccggtacctt aattaattaa ggtaccaggt aagtgtaccc aattcgccct atagtgagtc
     1501 gtattacaat tcactcgatc gcccttccca acagttgcgc agcctgaatg gcgaatggag
     1561 atccaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag
     1621 attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata
     1681 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1741 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     1801 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1861 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     1921 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1981 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2041 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2101 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2161 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2221 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2281 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2341 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2401 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2461 ataaatgctt caataatatt gaaaaaggaa gaatcctgag gcggaaagaa ccagctgtgg
     2521 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2581 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2641 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2701 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2761 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2821 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     2881 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     2941 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3001 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3061 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3121 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3181 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3241 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3301 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3361 ggacgaagaa catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3421 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3481 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3541 tcaggacata gcgttggcta cccgtgatat tgctgaagaa cttggcggcg aatgggctga
     3601 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3661 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3721 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3781 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3841 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     3901 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     3961 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4021 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4081 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4141 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4201 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4261 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4321 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4381 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4441 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4501 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4561 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4621 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4681 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4741 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4801 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4861 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4921 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4981 accgccatgc
//
