LOCUS       sequence-367393-        7344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human SOX2 and miRFP670 via T2A linker under control of a 
            CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7344)
  AUTHORS   Abatti LE, Lado-Fernandez P, Huynh L, Collado M, Hoffman MM, 
            Mitchell JA
  TITLE     Epigenetic reprogramming of a distal developmental enhancer cluster 
            drives SOX2 overexpression in breast and lung adenocarcinoma.
  JOURNAL   Nucleic Acids Res. 2023 Sep 22:gkad734. doi: 10.1093/nar/gkad734.
  PUBMED    37738673
REFERENCE   2  (bases 1 to 7344)
  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: "Nucleic 
            Acids Res. 2023 Sep 22:gkad734. doi: 10.1093/nar/gkad734."
COMMENT     Sequence Label: CMV-SOX2-T2A-miRFP670
FEATURES             Location/Qualifiers
     source          1..7344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        169..548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        549..752
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     703..723
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     749..773
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1808..1861
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1868..2812
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="VAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDHR
                     VIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTEY
                     CGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTGY
                     DRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVTY
                     QPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLVV
                     CHHYLPRFIRFELRAICKRLAERIATRITALES"
     primer_bind     complement(2849..2868)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2893..2941
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3143..3571
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3230..3249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3440..3461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3580..3600)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3585..3914
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3765..3900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3827..3846
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3981..4775
                     /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(4035..4054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4645..4664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4951..5072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4988..5007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5042..5061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5121..5137)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5121..5137)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5134..5156)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5145..5161
                     /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(5169..5175,5176..5193,5194..5199))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5214..5235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5352..5369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5523..6111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5603..5622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6282..7073,7074..7142))
                     /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     6905..6924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7143..7247)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7322..7341)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
       61 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      121 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      181 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      241 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      301 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      361 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      421 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      481 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      541 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      601 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      661 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      721 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
      781 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      841 ctctagaatg tacaacatga tggagacgga gctgaagccg ccgggcccgc agcaaacttc
      901 ggggggcggc ggcggcaact ccaccgcggc ggcggccggc ggcaaccaga aaaacagccc
      961 ggaccgcgtc aagcggccca tgaatgcctt catggtgtgg tcccgcgggc agcggcgcaa
     1021 gatggcccag gagaacccca agatgcacaa ctcggagatc agcaagcgcc tgggcgccga
     1081 gtggaaactt ttgtcggaga cggagaagcg gccgttcatc gacgaggcta agcggctgcg
     1141 agcgctgcac atgaaggagc acccggatta taaataccgg ccccggcgga aaaccaagac
     1201 gctcatgaag aaggataagt acacgctgcc cggcgggctg ctggcccccg gcggcaatag
     1261 catggcgagc ggggtcgggg tgggcgccgg cctgggcgcg ggcgtgaacc agcgcatgga
     1321 cagttacgcg cacatgaacg gctggagcaa cggcagctac agcatgatgc aggaccagct
     1381 gggctacccg cagcacccgg gcctcaatgc gcacggcgca gcgcagatgc agcccatgca
     1441 ccgctacgac gtgagcgccc tgcagtacaa ctccatgacc agctcgcaga cctacatgaa
     1501 cggctcgccc acctacagca tgtcctactc gcagcagggc acccctggca tggctcttgg
     1561 ctccatgggt tcggtggtca agtccgaggc cagctccagc ccccctgtgg ttacctcttc
     1621 ctcccactcc agggcgccct gccaggccgg ggacctccgg gacatgatca gcatgtatct
     1681 ccccggcgcc gaggtgccgg aacccgccgc ccccagcaga cttcacatgt cccagcacta
     1741 ccagagcggc ccggtgcccg gcacggccat taacggcaca ctgcccctct cacacatgtt
     1801 ggctagcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     1861 tggtaccgta gcaggtcatg cctctggcag ccccgcattc gggaccgcct ctcattcgaa
     1921 ttgcgaacat gaagagatcc acctcgccgg ctcgatccag ccgcatggcg cgcttctggt
     1981 cgtcagcgaa catgatcatc gcgtcatcca ggccagcgcc aacgccgcgg aatttctgaa
     2041 tctcggaagc gtactcggcg ttccgctcgc cgagatcgac ggcgatctgt tgatcaagat
     2101 cctgccgcat ctcgatccca ccgccgaagg catgccggtc gcggtgcgct gccggatcgg
     2161 caatccctct acggagtact gcggtctgat gcatcggcct ccggaaggcg ggctgatcat
     2221 cgaactcgaa cgtgccggcc cgtcgatcga tctgtcaggc acgctggcgc cggcgctgga
     2281 gcggatccgc acggcgggtt cactgcgcgc gctgtgcgat gacaccgtgc tgctgtttca
     2341 gcagtgcacc ggctacgacc gggtgatggt gtatcgtttc gatgagcaag gccacggcct
     2401 ggtattctcc gagtgccatg tgcctgggct cgaatcctat ttcggcaacc gctatccgtc
     2461 gtcgactgtc ccgcagatgg cgcggcagct gtacgtgcgg cagcgcgtcc gcgtgctggt
     2521 cgacgtcacc tatcagccgg tgccgctgga gccgcggctg tcgccgctga ccgggcgcga
     2581 tctcgacatg tcgggctgct tcctgcgctc gatgtcgccg tgccatctgc agttcctgaa
     2641 ggacatgggc gtgcgcgcca ccctggcggt gtcgctggtg gtcggcggca agctgtgggg
     2701 cctggttgtc tgtcaccatt atctgccgcg cttcatccgt ttcgagctgc gggcgatctg
     2761 caaacggctc gccgaaagga tcgcgacgcg gatcaccgcg cttgagagct aaaccggtta
     2821 gtaatgagtt taaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2881 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     2941 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3001 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3061 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3121 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3181 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3241 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3301 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3361 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3421 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3481 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3541 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3601 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3661 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3721 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3781 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3841 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3901 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3961 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4021 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4081 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4141 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4201 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4261 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4321 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4381 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4441 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4501 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4561 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4621 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4681 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4741 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4801 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4861 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4921 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4981 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5041 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5101 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5161 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5221 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5281 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5341 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5401 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5461 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5521 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5581 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5641 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5701 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5761 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5821 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5881 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5941 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6001 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6061 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6121 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6181 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6241 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6301 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6361 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6421 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6481 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6541 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6601 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6661 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6721 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6781 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6841 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6901 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6961 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7021 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7081 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7141 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7201 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7261 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     7321 gtacaatctg ctctgatgcc gcat
//
