LOCUS       sequence-166161-        7583 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 7583)
  AUTHORS   Maruyama M, Ichisaka T, Nakagawa M, Yamanaka S
  TITLE     Differential roles for Sox15 and Sox2 in transcriptional control in 
            mouse embryonic stem cells.
  JOURNAL   J Biol Chem. 2005 Jul 1. 280(26):24371-9.
  PUBMED    15863505
REFERENCE   2  (bases 1 to 7583)
  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: "J Biol 
            Chem. 2005 Jul 1. 280(26):24371-9."
COMMENT     Sequence Label: pCAG-HA-Sox2-IP
FEATURES             Location/Qualifiers
     source          1..7583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1743..1769
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1779..1803
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2764..2788)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(2936..3510,3511..3513,3514..3522)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3103..3120)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3330..3349
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3523..4122
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3523..3542)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4019..4039
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4203..4220)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4209..4433
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4480..4496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4480..4496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4493..4515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4504..4520
                     /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(4528..4534,4535..4552,4553..4558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4573..4594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4653..4848
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4699..4834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4854..4988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5056..5073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5227..5815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5307..5326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5986..6777,6778..6846))
                     /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     6609..6628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7439..7573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcaccatgg
     1741 ggtacccata cgatgttccg gattacgctg gatcatcaac aagtttgtac aaaaaagcag
     1801 gcttgtataa catgatggag acggagctga agccgccggg cccgcagcaa gcttcggggg
     1861 gcggcggcgg aggaggcaac gccacggcgg cggcgaccgg cggcaaccag aagaacagcc
     1921 cggaccgcgt caagaggccc atgaacgcct tcatggtatg gtcccggggg cagcggcgta
     1981 agatggccca ggagaacccc aagatgcaca actcggagat cagcaagcgc ctgggcgcgg
     2041 agtggaaact tttgtccgag accgagaagc ggccgttcat cgacgaggcc aagcggctgc
     2101 gcgctctgca catgaaggag cacccggatt ataaataccg gccgcggcgg aaaaccaaga
     2161 cgctcatgaa gaaggataag tacacgcttc ccggaggctt gctggccccc ggcgggaaca
     2221 gcatggcgag cggggttggg gtgggcgccg gcctgggtgc gggcgtgaac cagcgcatgg
     2281 acagctacgc gcacatgaac ggctggagca acggcagcta cagcatgatg caggagcagc
     2341 tgggctaccc gcagcacccg ggcctcaacg ctcacggcgc ggcacagatg caaccgatgc
     2401 accgctacga cgtcagcgcc ctgcagtaca actccatgac cagctcgcag acctacatga
     2461 acggctcgcc cacctacagc atgtcctact cgcagcaggg cacccccggt atggcgctgg
     2521 gctccatggg ctctgtggtc aagtccgagg ccagctccag cccccccgtg gttacctctt
     2581 cctcccactc cagggcgccc tgccaggccg gggacctccg ggacatgatc agcatgtacc
     2641 tccccggcgc cgaggtgccg gagcccgctg cgcccagtag actgcacatg gcccagcact
     2701 accagagcgg cccggtgccc ggcacggcca ttaacggcac actgcccctg tcgcacatgt
     2761 gaaacccagc tttcttgtac aaagtggttg atgatccctc gaggaattca ctcctcaggt
     2821 gcaggctgcc tatcagaagg tggtggctgg tgtggccaac tggctcacaa ataccactga
     2881 gatcgacggt atcgagcggc cgctcgataa gcttgatatc gaattccgcc ccccccccct
     2941 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     3001 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     3061 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     3121 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     3181 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     3241 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     3301 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     3361 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3421 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3481 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatgaccga gtacaagccc
     3541 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3601 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3661 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3721 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3781 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3841 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3901 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3961 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4021 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4081 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     4141 cagcgcccga ccgaaaggag cgcacgaccc catgcatcga tgatctagag ctcgctgatc
     4201 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4261 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4321 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4381 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4441 ggcggaaaga acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4621 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     4681 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4741 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4801 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     4861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4981 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5041 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5101 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5161 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5221 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5281 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5341 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5401 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5461 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5521 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5581 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5641 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5701 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5761 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5821 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5881 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5941 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6001 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6061 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6121 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6181 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6241 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6301 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6361 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6421 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6481 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6541 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6601 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6661 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6721 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6781 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6841 actcatactc ttcctttttc aatgaccgct ttggaaaaac aaagactgta tttcctggaa
     6901 attaatgttt attcaataaa ctgtgtattc agctatattc acatagtggt gaggctgaaa
     6961 tgaggcggga agaggcggtt ggggcttaat tatatcaatt tgggtggccc cacagcgcct
     7021 ccaaggcgcc agtcctgttt tgacaagttg cctctggaag cctctcttct ttttctccag
     7081 agtaagcgga ggccaggggc ccccggcctc tgcttaatac taaaaaaaac agctgttgtc
     7141 atagtaatga ttgggtggaa acattccagg cctgggtgga gaggcttttt gcttcctctt
     7201 gcaaaaccac actgacattc caggcctggg tggagaggct ttttgcttcc tcttgcaaaa
     7261 ccacactgcc ctctggaggg cagttgccta gcaactaatt aaaagaggat gtcgcacggc
     7321 cagctgcggt cagttagtca cttcctgctt aactgacttg acattttcta ttttaagagt
     7381 cgggaggaaa attactgtgt tggaggccct ccgccatctt ctgaagctga atcgaattaa
     7441 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     7501 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7561 tcatgtctgg atctgatatc atc
//
