LOCUS       Exported                7040 bp DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Luciferase-pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  AUTHORS   Safran M, Kim WY, O'Connell F, Flippin L, Gunzler V, Horner JW, 
            Depinho RA, Kaelin WG
  TITLE     Mouse model for noninvasive imaging of HIF prolyl hydroxylase 
            activity: assessment of an oral agent that stimulates erythropoietin
            production.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Jan 3. 103(1):105-10.
  PUBMED    16373502
REFERENCE   2  (bases 1 to 7040)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2006 Jan 3. 103(1):105-10."
FEATURES             Location/Qualifiers
     source          1..7040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(105..125)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        110..439
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      290..425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     352..371
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             506..1300
                     /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(560..579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1170..1189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1474..1595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1511..1530)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1565..1584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1644..1660)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1644..1660)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1657..1679)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1668..1684
                     /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(1692..1722)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1737..1758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1875..1892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2046..2634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2126..2145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2805..3665)
                     /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     3428..3447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3666..3770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3845..3864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4036..4415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4416..4619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4570..4590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4664..4683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4664..4682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4719..4728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4725..6377
                     /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(4791..4809)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(6394..6412)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6395..6412)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6432..6449)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6438..6662
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6708..7040,1..96)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6795..6814)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7005..7026
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
       61 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
      121 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      181 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      241 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
      301 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
      361 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
      421 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
      481 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
      541 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
      601 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
      661 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
      721 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
      781 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
      841 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
      901 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
      961 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     1021 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     1081 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     1141 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     1201 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1261 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     1321 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     1381 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     1441 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     1501 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1561 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     1621 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1681 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1741 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1801 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1861 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1921 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1981 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2041 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2101 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2161 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2221 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2281 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2341 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2401 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2461 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2521 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2581 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2641 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2701 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2761 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2821 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2881 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2941 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3001 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3061 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3121 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3181 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3241 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3301 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3361 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3421 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3481 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3541 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3601 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3661 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3721 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3781 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     3841 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     3901 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     3961 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     4021 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     4081 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4141 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4201 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4261 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4321 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4381 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     4441 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4501 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4561 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     4621 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     4681 gggagaccca agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa
     4741 acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac
     4801 tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac
     4861 atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag
     4921 ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc
     4981 ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga
     5041 acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg
     5101 tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca
     5161 tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca
     5221 cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct
     5281 tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta
     5341 aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta
     5401 tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg
     5461 gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt
     5521 atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc
     5581 tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt
     5641 tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag
     5701 cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta
     5761 catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg
     5821 ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc
     5881 aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg
     5941 aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact
     6001 gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag
     6061 gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg
     6121 acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg
     6181 ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag
     6241 taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc
     6301 ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg
     6361 gaaagatcgc cgtgtaattc tagagggccc tattctatag tgtcacctaa atgctagagc
     6421 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     6481 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     6541 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6601 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     6661 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     6721 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6781 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6841 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     6901 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     6961 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7021 aactggaaca acactcaacc
//
