LOCUS       sequence-152187-        7298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of human Six1 and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7298)
  TITLE     Heide Ford plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7298)
  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: 
            "Unpublished"
COMMENT     Sequence Label: MSCV-Six1
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             36..552
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    616..957
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1024..1440
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1370..1392
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1408..1424
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2326..2893
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2343..2895
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2487..2504)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2714..2733
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2907..2909,2910..2912,2913..3626)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2952..2973)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3213..3232)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3560..3581
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3728..4242
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4404..4420
                     /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(4428..4434,4435..4452,4453..4458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4473..4494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4611..4628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4782..5370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4862..4881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5541..6332,6333..6401))
                     /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     6164..6183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6402..6506)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6574..6592
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6630..6652)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6752..6771
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6965..6987
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7115..7134
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
       61 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
      121 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
      181 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
      241 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
      301 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
      361 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
      421 gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg
      481 aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg
      541 gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc
      601 cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt
      661 gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt
      721 atctggcgga cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga
      781 cgtcccaggg actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg
      841 aatccgaccc cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg
      901 cctccgtctg aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag
      961 cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat
     1021 tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
     1081 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
     1141 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
     1201 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
     1261 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
     1321 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
     1381 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
     1441 ggaattagat catgtcgatg ctgccgtcgt ttggctttac gcaggagcaa gtggcgtgcg
     1501 tgtgcgaggt tctgcagcaa ggcggaaacc tggagcgcct gggcaggttc ctgtggtcac
     1561 tgcccgcctg cgaccacctg cacaagaacg agagcgtact caaggccaag gcggtggtcg
     1621 ccttccaccg cggcaacttc cgtgagctct acaagatcct ggagagccac cagttctcgc
     1681 ctcacaacca ccccaaactg cagcaactgt ggctgaaggc gcattacgtg gaggccgaga
     1741 agctgcgcgg ccgacccctg ggcgccgtgg gcaaatatcg ggtgcgccga aaatttccac
     1801 tgccgcgcac catctgggac ggcgaggaga ccagctactg cttcaaggag aagtcgaggg
     1861 gtgtcctgcg ggagtggtac gcgcacaatc cctacccatc gccgcgtgag aagcgggagc
     1921 tggccgaggc caccggcctc accaccaccc aggtcagcaa ctggtttaag aaccggaggc
     1981 aaagagaccg ggccgcggag gccaaggaaa gggagaacac cgaaaacaat aactcctcct
     2041 ccaacaagca gaaccaactc tctcctctgg aagggggcaa gccgctcatg tccagctcag
     2101 aagaggaatt ctcacctccc caaagtccag accagaactc agtccttctg ctgcagggca
     2161 atatgggcca cgccaggagc tcaaactatt ctctcccggg cttaacagcc tcgcagccca
     2221 gtcacggcct gcagacccac cagcatcagc tccaagactc tctgctcggc cccctcacct
     2281 ccagtctggt ggacttgggg tcctaagtgc tcgaggttaa cgaattccgc cccccccccc
     2341 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2401 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2461 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2521 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2581 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2641 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2701 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2761 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     2821 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     2881 aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     2941 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtctg
     3001 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3061 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3121 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3181 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3241 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3301 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3361 atatcatggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca
     3421 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3481 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     3541 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3601 tcggcatgga cgagctgtac aagtaatgaa ttaattaaga attatcaagc ttatcgatac
     3661 cgtcgacctg cagccaagct tatcgataaa ataaaagatt ttatttagtc tccagaaaaa
     3721 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     3781 caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag
     3841 agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     3901 ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc
     3961 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     4021 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     4081 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt
     4141 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     4201 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa
     4261 gttggagaac aacattctga gggtaggagt cgaatattaa gtaatcctga ctcaattagc
     4321 cactgttttg aatccacata ctccaatact cctgaaatag ttcattatgg acagcgcaga
     4381 aagagctggg gagaattgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4441 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4501 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4561 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4621 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4681 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4741 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4801 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4861 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4921 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4981 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5041 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5101 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5161 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5221 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5281 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5341 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5401 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5461 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5521 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5581 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5641 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5701 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5761 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5821 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5881 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5941 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6001 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6061 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6121 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6181 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6241 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6301 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6361 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6421 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6481 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6541 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6601 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6661 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6721 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6781 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6841 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6901 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6961 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
     7021 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     7081 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     7141 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
     7201 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     7261 accagctgaa gcctatagag tacgagccat agataaaa
//
