LOCUS       Exported                6709 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    2X PRE TK luc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  AUTHORS   Giangrande PH, Kimbrel EA, Edwards DP, McDonnell DP
  TITLE     The opposing transcriptional activities of the two isoforms of the 
            human progesterone receptor are due to differential cofactor 
            binding.
  JOURNAL   Mol Cell Biol. 2000 May . 20(9):3102-15.
  PUBMED    10757795
REFERENCE   2  (bases 1 to 6709)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6709
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(943..1803)
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1976..1994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2032..2054)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2154..2173
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2655..3168)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2850..2871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3062..3081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3594..3616
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3608..3625
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3609..3625
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3794..3856
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             3916..5568
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(3982..4000)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          5699..5764
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5894..5914
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6339..6473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6376..6395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6430..6449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6491..6507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6491..6507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6504..6526)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6515..6531
                     /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(6539..6569)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6584..6605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1981 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2041 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2101 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2161 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2221 gagaaaatac cgcatcaggc gattccaaca tccaataaat catacaggca aggcaaagaa
     2281 ttagcaaaat taagcaataa agcctcagag cataaagcta aatcggttgt accaaaaaca
     2341 ttatgaccct gtaatacttt tgcgggagaa gcctttattt caacgcaagg ataaaaattt
     2401 ttagaaccct catatatttt aaatgcaatg cctgagtaat gtgtaggtaa agattcaaac
     2461 gggtgagaaa ggccggagac agtcaaatca ccatcaatat gatattcaac cgttctagct
     2521 gataaattca tgccggagag ggtagctatt tttgagaggt ctctacaaag gctatcaggt
     2581 cattgcctga gagtctggag caaacaagag aatcgatgaa cggtaatcgt aaaactagca
     2641 tgtcaatcat atgtaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
     2701 aagcaaatat ttaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2761 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2821 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2881 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2941 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3001 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3061 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3121 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct
     3181 ttgacgagca cgtataacgt gctttcctcg ttagaatcag agcgggagct aaacaggagg
     3241 ccgattaaag ggattttaga caggaacggt acgccagaat cctgagaagt gtttttataa
     3301 tcagtgaggc caccgagtaa aagagtctgt ccatcacgca aattaaccgt tgtcgcaata
     3361 cttctttgat tagtaataac atcacttgcc tgagtagaag aactcaaact atcggccttg
     3421 ctggtaatat ccagaacaat attaccgcca gccattgcaa cggaatcgcc attcgccatt
     3481 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     3541 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     3601 acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg actctagagg
     3661 atccgtagct agaacatcct gtacaggatc cgtagctaga acatcctgta caggatccgg
     3721 ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg
     3781 gtccgaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct
     3841 gcagcgaccc gcttaacagc gtcaacagcg tgccgcagat ctcgaggagc ttggcattcc
     3901 ggtactgttg gtaaaatgga agacgccaaa aacataaaga aaggcccggc gccattctat
     3961 cctctagagg atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg
     4021 gttcctggaa caattgcttt tacagatgca catatcgagg tgaacatcac gtacgcggaa
     4081 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat
     4141 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg
     4201 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc
     4261 aacagtatga acatttcgca gcctaccgta gtgtttgttt ccaaaaaggg gttgcaaaaa
     4321 attttgaacg tgcaaaaaaa attaccaata atccagaaaa ttattatcat ggattctaaa
     4381 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt
     4441 tttaatgaat acgattttgt accagagtcc tttgatcgtg acaaaacaat tgcactgata
     4501 atgaattcct ctggatctac tgggttacct aagggtgtgg cccttccgca tagaactgcc
     4561 tgcgtcagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact
     4621 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat
     4681 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttttacga
     4741 tcccttcagg attacaaaat tcaaagtgcg ttgctagtac caaccctatt ttcattcttc
     4801 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggg
     4861 ggcgcacctc tttcgaaaga agtcggggaa gcggttgcaa aacgcttcca tcttccaggg
     4921 atacgacaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg
     4981 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat
     5041 ctggataccg ggaaaacgct gggcgttaat cagagaggcg aattatgtgt cagaggacct
     5101 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat
     5161 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatagtt
     5221 gaccgcttga agtctttaat taaatacaaa ggatatcagg tggcccccgc tgaattggaa
     5281 tcgatattgt tacaacaccc caacatcttc gacgcgggcg tggcaggtct tcccgacgat
     5341 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa
     5401 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga
     5461 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc
     5521 agagagatcc tcataaaggc caagaagggc ggaaagtcca aattgtaaaa tgtaactgta
     5581 ttcagcgatg acgaaattct tagctattgt aatactctag aggatctttg tgaaggaacc
     5641 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
     5701 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
     5761 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga
     5821 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca
     5881 acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact ttccttcaga
     5941 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat
     6001 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt
     6061 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca
     6121 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt
     6181 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa
     6241 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     6301 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     6361 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6421 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcgggaatt
     6481 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6541 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6601 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6661 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcg
//
