LOCUS       Exported                4923 bp ds-DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pREBFB15 = pGL4-XX [luc2P-(GAS/ISRE [STAT1/STAT1]; 4x direct 
            repeats)-Hygro] created by Brock F. Binkowski

            

            Spe1/Xho1 double digest on syhthetic construct 'HSE-1' from IDT and 
            pGL4.27(buffer B);

            ligate and transform.

            

            Failed attempt: Amplify pGL4.27 w/ BFB861/876;

            Spe1/Xho1 double digest (buffer B)of purified PCR product and 
            pGL4.27;

            ligate and transform JM109
FEATURES             Location/Qualifiers
     source          1..4923
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    27..110
                     /label=GAS-RE
                     /note="GAS-RE"
     promoter        128..158
                     /label=minimal promoter
                     /note="minimal promoter"
     misc_feature    191..829
                     /label=NanoLucP
                     /note="NanoLucP"
     polyA_signal    869..1090
                     /label=SV40 late poly(A) signal
                     /note="SV40 late poly(A) signal"
     misc_feature    1138..1556
                     /label=SV40 promoter
                     /note="SV40 promoter"
     CDS             1581..2618
                     /codon_start=1
                     /label=HygroR
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     rep_origin      3014
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="ColE1-derived plasmid replication origin"
     gene            complement(3805..4665)
                     /label=AmpR
     polyA_signal    4770..4923
                     /label=poly(A)  pause site
                     /note="poly(A)  pause site"
ORIGIN
        1 ggcctaactg gccggtacct gagctcagtt tcatattact ctaaatcagt ttcatattac
       61 tctaaatcag tttcatatta ctctaaatca gtttcatatt actctaaatc ctcgaagctt
      121 agacactaga gggtatataa tggaagctcg acttccagct tggcaatccg gtactgttgg
      181 taaagccacc atggtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg
      241 ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg
      301 ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga
      361 catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa
      421 aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg
      481 cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga
      541 aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa
      601 caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat
      661 caacggagtg accggctggc ggctgtgcga acgcattctg gcgaattctc acggctttcc
      721 gcctgaggtt gaagagcaag ccgccggtac attgcctatg tcctgcgcac aagaaagcgg
      781 tatggaccgg cacccagccg cttgtgcttc agctcgcatc aacgtctaag gccgcgactc
      841 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
      901 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      961 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1021 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1081 aaatgtggta aaatcgataa ggatccgttt gcgtattggg cgctcttccg ctgatctgcg
     1141 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttagctacc ttctgaggcg
     1201 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1381 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1441 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1501 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgatt
     1561 cttctgacac tagcgccacc atgaagaagc ccgaactcac cgctaccagc gttgaaaaat
     1621 ttctcatcga gaagttcgac agtgtgagcg acctgatgca gttgtcggag ggcgaagaga
     1681 gccgagcctt cagcttcgat gtcggcggac gcggctatgt actgcgggtg aatagctgcg
     1741 ctgatggctt ctacaaagac cgctacgtgt accgccactt cgccagcgct gcactaccca
     1801 tccccgaagt gttggacatc ggcgagttca gcgagagcct gacatactgc atcagtagac
     1861 gcgcccaagg cgttactctc caagacctcc ccgaaacaga gctgcctgct gtgttacagc
     1921 ctgtcgccga agctatggat gctattgccg ccgccgacct cagtcaaacc agcggcttcg
     1981 gcccattcgg gccccaaggc atcggccagt acacaacctg gcgggatttc atttgcgcca
     2041 ttgctgatcc ccatgtctac cactggcaga ccgtgatgga cgacaccgtg tccgccagcg
     2101 tagctcaagc cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag gtgcgccacc
     2161 tcgtccatgc cgacttcggc agcaacaacg tcctgaccga caacggccgc atcaccgccg
     2221 taatcgactg gtccgaagct atgttcgggg acagtcagta cgaggtggcc aacatcttct
     2281 tctggcggcc ctggctggct tgcatggagc agcagactcg ctacttcgag cgccggcatc
     2341 ccgagctggc cggcagccct cgtctgcgag cctacatgct gcgcatcggc ctggatcagc
     2401 tctaccagag cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa ggccgctgcg
     2461 atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct cgccggagcg
     2521 cagccgtatg gaccgacggc tgcgtcgagg tgctggccga cagcggcaac cgccggccca
     2581 gtacacgacc gcgcgctaag gaggtaggtc gagtttaaac tctagaaccg gtcatggccg
     2641 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg ttcgaactag
     2701 atgctgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     2761 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     2821 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc
     3781 gcaaatgcta aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc
     3841 gatctgccta tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat
     3901 tcgtgagggc ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc
     3961 ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc
     4021 tgctactttg tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag
     4081 ttcgccagtg agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg
     4141 ctcgtcgttc ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg
     4201 atcacccata ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag
     4261 taagttggcc gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt
     4321 catgccatcc gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga
     4381 gtagtgtata cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc
     4441 acatagcagt actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc
     4501 aaggatcttg ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc
     4561 ttcagcatct tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc
     4621 cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca
     4681 atattattga agcatttatc agggttacta gtacgtctct caaggataag taagtaatat
     4741 taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt
     4801 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     4861 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     4921 tct
//
