LOCUS       sequence-345436-        7455 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 7455)
  AUTHORS   Shen WF, Montgomery JC, Rozenfeld S, Moskow JJ, Lawrence HJ, 
            Buchberg AM, Largman C
  TITLE     AbdB-like Hox proteins stabilize DNA binding by the Meis1 
            homeodomain proteins.
  JOURNAL   Mol Cell Biol 1997 Nov;17(11):6448-58.
  PUBMED    9343407
REFERENCE   2  (bases 1 to 7455)
  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"; date: "1997-11"; volume: "17(11)"; pages: "6448-58"
COMMENT     Sequence Label: HOXA9 (murine) FLAG MSCV
FEATURES             Location/Qualifiers
     source          1..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..342
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             409..825
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     755..777
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     793..809
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             837..860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2003..2464
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2059..2076)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2286..2305
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2469..2471,2472..2474,2475..3188)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2469..2471,2472..2474,2475..3185)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2514..2535)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2775..2794)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3122..3143
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3270..3784
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3946..3962
                     /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(3970..3976,3977..3994,3995..4000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4015..4036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4153..4170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4324..4912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4404..4423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5083..5874,5875..5943))
                     /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     5706..5725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5944..6048)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6116..6134
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6172..6194)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6294..6313
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6507..6529
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6657..6676
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6876..7392
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc
       61 taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt
      121 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
      181 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
      241 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
      301 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
      361 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
      421 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
      481 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
      541 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
      601 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
      661 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
      721 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
      781 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat tctatggact
      841 acaaggacga cgatgacaag cttaagaggc ctcaggatcc caccaccggg gccctgggca
      901 actactatgt ggactccttc ctgctgggcg ccgacgctgc tgatgagctg ggtgcgggac
      961 gctacgctcc agggaccctg ggtcaacccc caaggcaggc ggcagctctg gccgaacacc
     1021 ccgacttcag tccttgcagc ttccagtcca aggcggcggt gtttggtgcc tcgtggaacc
     1081 cagtgcacgc ggcgggcgcc aatgcggtgc ctgctgcagt gtatcatcac caccaccacc
     1141 cctacgtgca tccccaggcg cccgtggcgg cggcggcgcc ggacggcagg tatatgcgct
     1201 cctggctgga acccacgccc ggtgcgctct ccttcgcggg cttaccctcc agccggcctt
     1261 atggcattaa acctgaaccg ctctcggcca gaaggggtga ctgtcccacg cttgacactc
     1321 acactttgtc cctgactgac tatgcttgtg gttctcctcc agttgataga gaaaaacaac
     1381 ccagcgaagg cgccttctcc gaaaacaatg ccgagaatga gagcggcgga gacaagcccc
     1441 ccatcgatcc caataacccg gctgccaact ggctacatgc tcgctccact cggaagaagc
     1501 gatgccccta cacaaaacac cagacgctgg aactggagaa ggagtttctg tttaacatgt
     1561 acctcacacg ggaccgcagg tacgaggtgg cccggctgct caacctcacc gaaaggcagg
     1621 tcaagatctg gttccagaac cgcaggatga aaatgaagaa aatcaacaag gaccgagcaa
     1681 aagacgagtg agccttttag gggctcattt aaaaagagag caagctagaa agaaaaagaa
     1741 aggactgtcc gtctccctct gtctcctctc ccccaaaccc agcctccacc cgcacaaagg
     1801 ggctctaaat cccaggcctc atctccccac tggcagtccg tgctcaggct ggctcttagg
     1861 cctgcggctt tgatggagga ggtattgtaa gcttggtacc gagctcggat ccactagtaa
     1921 cggccgccag tgtgctggaa ttcgttaacc tcgagaaaac tcgacggtta ttttccacca
     1981 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     2041 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2101 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     2161 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     2221 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     2281 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     2341 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     2401 taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     2461 ataataccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     2521 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     2581 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2641 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2701 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2761 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2821 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2881 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2941 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3001 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3061 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3121 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3181 acaagtaaag cggccaactc gacgttattc ccttcgaagg cctatcgata aaataaaaga
     3241 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     3301 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     3361 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     3421 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     3481 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     3541 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     3601 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     3661 tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     3721 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     3781 tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt
     3841 aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat
     3901 agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac
     3961 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4021 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4081 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4141 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4201 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4261 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4321 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4381 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4441 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4501 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4561 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4621 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4681 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4741 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4801 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4861 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4921 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4981 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5041 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5101 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5161 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5221 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5281 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5341 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5401 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5461 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5521 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5581 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5641 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5701 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5761 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5821 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5881 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5941 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6001 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6061 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6121 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6181 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6241 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6301 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6361 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6421 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6481 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg
     6541 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     6601 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     6661 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     6721 cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct
     6781 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     6841 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     6901 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     6961 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     7021 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     7081 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     7141 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     7201 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     7261 gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg
     7321 aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg
     7381 gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc
     7441 cccccgccgg gaggt
//
