LOCUS       Exported                5097 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGL3-RARE-luciferase
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5097)
  AUTHORS   Hoffman LM, Garcha K, Karamboulas K, Cowan MF, Drysdale LM, Horton 
            WA, Underhill TM
  TITLE     BMP action in skeletogenesis involves attenuation of retinoid 
            signaling.
  JOURNAL   J Cell Biol. 2006 Jul 3. 174(1):101-13.
  PUBMED    16818722
REFERENCE   2  (bases 1 to 5097)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5097
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        135..331
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      182..317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     244..263
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     regulatory      361..370
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             367..2019
                     /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(433..451)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2060..2181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2071..2090)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2125..2144
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2429..2446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2600..3188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2680..2699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3359..4219)
                     /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     3982..4001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4220..4324)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4351..4806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4438..4457)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4648..4669
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4937..4985
                     /note="synthetic polyadenylation signal"
     misc_feature    4999..5090
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5039..5058
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcggtgaact ttcggtgaac cctaccctcg agtgaacttt cggtgaaccc
       61 taccctcgag ggtagggttc accgaaagtt cactcgacga attccagtct agcccgggct
      121 cgagatctgc gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
      181 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatcgct gactaatttt
      241 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
      301 aggctttttt ggaggcctag gcttttgcaa aaagcttggc attccggtac tgttggtaaa
      361 gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa
      421 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga
      481 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa
      541 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc
      601 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc
      661 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg
      721 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac
      781 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac
      841 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa
      901 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc
      961 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga
     1021 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta
     1081 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt
     1141 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag
     1201 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc
     1261 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc
     1321 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa
     1381 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa
     1441 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc
     1501 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg
     1561 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta
     1621 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg
     1681 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg
     1741 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt
     1801 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc
     1861 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt
     1921 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc
     1981 ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat tctagagtcg gggcggccgg
     2041 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2101 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2161 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2221 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     2281 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     2341 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     2401 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2461 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2521 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2581 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2641 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2701 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2761 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2821 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2881 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2941 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3001 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3061 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3121 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3181 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3241 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3301 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3361 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3421 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3481 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3541 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3601 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3661 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3721 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3781 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3841 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3901 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3961 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4021 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4081 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4141 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4201 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4261 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4321 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     4381 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4441 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4501 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4561 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4621 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4681 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4741 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4801 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4861 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     4921 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4981 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     5041 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgata
//
