LOCUS       sequence-220060-        5502 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 5502)
  AUTHORS   Graves RA, Tontonoz P, Spiegelman BM
  TITLE     Analysis of a tissue-specific enhancer: ARF6 regulates adipogenic 
            gene expression.
  JOURNAL   Mol Cell Biol. 1992 Mar . 12(3):1202-8.
  PUBMED    1545801
REFERENCE   2  (bases 1 to 5502)
  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. 1992 Mar . 12(3):1202-8."
COMMENT     Sequence Label: aP2-luciferase
FEATURES             Location/Qualifiers
     source          1..5502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    57..105
                     /note="synthetic polyadenylation signal"
     misc_feature    119..210
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     159..178
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        757..953
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      804..939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      983..992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             989..2641
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1055..1073)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2682..2803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2693..2712)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2747..2766
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3051..3068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3222..3810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3302..3321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3981..4772,4773..4841))
                     /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     4604..4623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4842..4946)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4973..5428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5060..5079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5270..5291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata
       61 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa
      121 catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca
      181 gtgcaagtgc aggtgccaga acatttctct atcgataggt accgaattcc agcaggaatc
      241 aggtagctgg agaatcgcac agagccatgc ggattcttgg caagccatgc gacaaaggca
      301 gaaatgcaca tttcacccag agagaaggga ttgtagtcag caggaagtca ccacccagag
      361 agcaaatgga gttcccagat gcctgacatt tgccttctta ctggatcaga gttcactagt
      421 ggaagtgtca cagcccaaac actcccccaa aggctcagcc cttccttgcc ttgtaacaat
      481 caagccgctc ctggatgaac tgctccgccc tctgtctctt tggcagggtt ggagcccact
      541 gtggcctgag cgacttctat ggctcccttt ctgtgatttt catggtttct gagctctttt
      601 cccccgcttt atgattttct ctttttgtct ctctcttgct aaacctcctt cgtatatgcc
      661 ctctcaggtt tcatttctga atcatctact gtgaactatt cccattgttt gccagaagcc
      721 ccctggttct tccttctaga ctcgagatct gcgatctgca tctcaattag tcagcaacca
      781 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      841 cgccccatcg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
      901 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
      961 gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa agaaaggccc
     1021 ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa
     1081 gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat
     1141 cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg
     1201 gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc
     1261 ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga
     1321 acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa
     1381 ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat
     1441 catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca
     1501 tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac
     1561 aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc
     1621 tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat
     1681 cattccggat actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac
     1741 tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga
     1801 gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg tgccaaccct
     1861 attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga
     1921 aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt
     1981 ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat
     2041 tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc
     2101 gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg
     2161 tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt
     2221 gattgacaag gatggatggc tacattctgg agacatagct tactgggacg aagacgaaca
     2281 cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc
     2341 cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg
     2401 tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa
     2461 gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa
     2521 gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga
     2581 cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta
     2641 attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2701 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2761 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2821 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     2881 ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc
     2941 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     3001 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     3061 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3121 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3181 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3241 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3301 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3361 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3421 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3481 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3541 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3601 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3661 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3721 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     4981 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5041 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5101 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5161 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5221 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5281 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5341 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5401 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     5461 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc cc
//
