LOCUS       sequence-342541-        8354 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 8354)
  AUTHORS   Shen WF, Hu YL, Uttarwar L, Passegue E, Largman C
  TITLE     MicroRNA-126 regulates HOXA9 by binding to the homeobox.
  JOURNAL   Mol Cell Biol. 2008 Jul . 28(14):4609-19.
  PUBMED    18474618
REFERENCE   2  (bases 1 to 8354)
  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. 2008 Jul . 28(14):4609-19."
COMMENT     Sequence Label: HOXA9-MSCV-full-GFP
FEATURES             Location/Qualifiers
     source          1..8354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1417..1440
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        3436..3935
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3471..3494)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3868..3887
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3942..3951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3948..3950,3951..3953,3954..4667)
                     /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(3948..3950,3951..3953,3954..4664)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3993..4014)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4254..4273)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4601..4622
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4749..5263
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5425..5441
                     /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(5449..5455,5456..5473,5474..5479))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5494..5515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5632..5649)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5803..6391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5883..5902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6562..7353,7354..7422))
                     /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     7185..7204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7423..7527)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7595..7613
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7651..7673)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7773..7792
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7986..8008
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8136..8155
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tctatggact acaaggacga cgatgacaag
     1441 cttaagaggc ctcaggatcc caccaccggg gccctgggca actactatgt ggactccttc
     1501 ctgctgggcg ccgacgctgc tgatgagctg ggtgcgggac gctacgctcc agggaccctg
     1561 ggtcaacccc caaggcaggc ggcagctctg gccgaacacc ccgacttcag tccttgcagc
     1621 ttccagtcca aggcggcggt gtttggtgcc tcgtggaacc cagtgcacgc ggcgggcgcc
     1681 aatgcggtgc ctgctgcagt gtatcatcac caccaccacc cctacgtgca tccccaggcg
     1741 cccgtggcgg cggcggcgcc ggacggcagg tatatgcgct cctggctgga acccacgccc
     1801 ggtgcgctct ccttcgcggg cttaccctcc agccggcctt atggcattaa acctgaaccg
     1861 ctctcggcca gaaggggtga ctgtcccacg cttgacactc acactttgtc cctgactgac
     1921 tatgcttgtg gttctcctcc agttgataga gaaaaacaac ccagcgaagg cgccttctcc
     1981 gaaaacaatg ccgagaatga gagcggcgga gacaagcccc ccatcgatcc caataacccg
     2041 gctgccaact ggctacatgc tcgctccact cggaagaagc gatgccccta cacaaaacac
     2101 cagacgctgg aactggagaa ggagtttctg tttaacatgt acctcacacg ggaccgcagg
     2161 tacgaggtgg cccggctgct caacctcacc gaaaggcagg tcaagatctg gttccagaac
     2221 cgcaggatga aaatgaagaa aatcaacaag gaccgagcaa aagacgagtg agccttttag
     2281 gggcacattt aaaaagagag caagctagaa agaaaaagaa aggactgtcc gtctccctct
     2341 gtctcctctc ccccaaaccc agcctccacc cgcacaaagg ggctctaaat cccaggcctc
     2401 atctccccac tggcagcccg tgctcaggct ggctcttagg cctgcggctt tgatggagga
     2461 ggtattgtaa gctttccatt ttatagaagg cacacacaca cacaagggag ggcattagcg
     2521 ctattggctg tatgtgctag cttgtatata tatatatata tttaaaaaaa atctacctgc
     2581 ttctgacttt aagcaaaagg aaagaaaact acctttttat ataatgcaca actgttgatg
     2641 actggctgta tagtttttag tctctgtagc taatttaatt tgctcttcgt gtggcagatc
     2701 attctgccaa aatacttgaa cactgtgttt tattgtggtg attatgtttt gtgactcaaa
     2761 cttctgtgct gggtgaagta cccattgtga ttgtggaagc tagaattcaa tttgaactca
     2821 ggttgtttat ggggggagag aagctgtggc ataatgtata gctctgtagt cttctgtgta
     2881 gctagactgt taacctacga ttgtaaacta gctgtaagga tttcccagtg aaatcagatt
     2941 tggaaagaaa agagagatag ttctccggga tgcatagatt catgatttat ctacttttga
     3001 aatgtgggct tcttcatctt tacgttctag actccaattt tgtccaatgt atatagaatc
     3061 tccatattaa agaaatattc ataatcagaa agcttgaata ttgtttttgc accagaggag
     3121 aagtgaggaa attcggagct atacgtgtgt gcagagggtt gctacctatg ggtttacgct
     3181 taattttaat tgggagaaag tgagtgctta ttgtaatgga gttaatttta ttgatgtacc
     3241 ttatacacca tgaaactgct cttgggctac tgtagatttg tatccttggt gaatttgggg
     3301 ttcccatcgg attgaacata cactgtatat tttgcaatag ttacctcaag gcctactgac
     3361 caaattgttg tgtcgagata tttaactttt gccaaacgga atatattgat tcttctcgag
     3421 agatctaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt
     3481 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca
     3541 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta
     3601 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac
     3661 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga
     3721 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc
     3781 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg
     3841 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg
     3901 cgctgttctc ctcttcctca tctccgggcc tttcggctgc agccaccatg gtgagcaagg
     3961 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     4021 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     4081 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     4141 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     4201 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     4261 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4321 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4381 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     4441 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     4501 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     4561 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     4621 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccaactcg
     4681 acgttattcc cttcgaaggc ctatcgataa aataaaagat tttatttagt ctccagaaaa
     4741 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     4801 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     4861 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     4921 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     4981 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     5041 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     5101 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg
     5161 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     5221 tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga
     5281 agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag
     5341 ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag
     5401 aaagagctgg ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5461 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5521 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5581 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5641 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5701 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5761 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5821 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5881 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5941 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6001 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6061 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6241 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6421 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6481 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6541 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6601 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6661 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6721 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6781 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6841 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6901 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6961 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7021 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7081 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7141 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7201 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7261 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7321 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7381 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7441 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7501 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     7561 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     7621 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     7681 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     7741 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     7801 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     7861 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     7921 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     7981 gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg
     8041 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     8101 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     8161 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta
     8221 gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat
     8281 caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc
     8341 cagaaaaagg gggg
//
