LOCUS       sequence-346546-        6456 bp    DNA     circular SYN 15-APR-2025
DEFINITION  This is a dual function destination vector for eSTARR-seq and 
            luciferase assay to quantify enhancer activity. It contains a MYC 
            promoter, a luciferase CDS, followed by a Gateway cassette (R2-R1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  TITLE     Transcription imparts architecture, function, and logic to enhancer 
            units
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6456)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pDEST-hSTARR-luc-Pmyc-ccw
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(120..125,126..143,144..150)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             204..884
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(270..289)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1204..1509
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1388..1407
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1550..1674)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1707..3359)
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3403..3422)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3405..3634
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     misc_feature    3925..4016
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3957..3976)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4030..4078
                     /note="synthetic polyadenylation signal"
     CDS             join(4183..4251,4252..5043)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5243..5831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5732..5751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5985..6002
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6247..6368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6284..6303)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6338..6357
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    join(6427..6456,1..95)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
       61 taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct
      121 ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat
      181 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
      241 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
      301 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
      361 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
      421 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
      481 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
      541 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
      601 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
      661 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
      721 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
      781 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
      841 atgagtggca ggcggggcgt aatctagagg atccggctta ctaaaagcca gataacagta
      901 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
      961 tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc
     1021 tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc
     1081 agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg
     1141 ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt
     1201 gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     1261 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
     1321 cgtctgctgt cagataaagt cccccgtgaa ctttacccgg tggtgcatat cggggatgaa
     1381 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     1441 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     1501 ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc atagtgactg
     1561 gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat
     1621 attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag tggtttgatc
     1681 aagctatcac cggtgcatgc tctagattac acggcgatct tgccgccctt cttggcctta
     1741 atgagaatct cgcggatctt gcgggcgtcc aacttgccgg tcagtccttt aggcacctcg
     1801 tccacgaaca caacaccacc gcgcagcttc ttggcggttg taacctggct ggccacatag
     1861 tccacgatct ccttctcggt catggtttta ccgtgttcca gcacgacgac tgcggcgggc
     1921 agctcgccgg catcgtcgtc gggcaggccg gcgaccccgg cgtcgaagat gttggggtgt
     1981 tgcagcagga tgctctccag ttcggctggg gctacctggt agcccttgta tttgatcagg
     2041 ctcttcagcc ggtccacgat gaagaagtgc tcgtcctcgt cccagtaggc gatgtcgccg
     2101 ctgtgcagcc agccgtcctt gtcgatgaga gcgtttgtag cctcggggtt gttaacgtag
     2161 ccgctcatga tcatggggcc acggacgcac agctcgccgc gctggttcac acccagtgtc
     2221 ttaccggtgt ccaagtccac caccttagcc tcgaagaagg gcaccacctt gcctactgcg
     2281 ccaggcttgt cgtccccttc gggggtgatc agaatggcgc tggttgtttc tgtcaggccg
     2341 tagccctggc ggatgcctgg taggtggaag cgtttggcca cggcctcacc tacctccttg
     2401 ctgagcggcg ccccgccgct ggcgatctcg tgcaagttgc ttaggtcgta cttgtcgatg
     2461 agagtgctct tagcgaagaa gctaaatagt gtgggcacca gcagggcaga ttgaatctta
     2521 tagtcttgca agctgcgcaa gaatagctcc tcctcgaagc ggtacatgag cacgacccga
     2581 aagccgcaga tcaagtagcc cagcgtggtg aacatgccga agccgtggtg aaatggcacc
     2641 acgctgagga tagcggtgtc ggggatgatc tggttgccga agatggggtc gcgggcatga
     2701 ctgaatcgga cacaagcggt gcggtgcggt agggctacgc ccttgggcaa tccggtactg
     2761 ccactactgt tcatgatcag ggcgatggtt ttgtcccggt cgaagctctc gggcacgaag
     2821 tcgtactcgt tgaagccggg tggcaaatgg gaagtcacga aggtgtacat gctttggaag
     2881 ccctggtagt cggtcttgct atccatgatg atgatctttt gtatgatcgg tagcttcttt
     2941 tgcacgttga ggatcttttg cagccctttc ttgctcacga atacgacggt gggctggctg
     3001 atgcccatgc tgttcagcag ctcgcgctcg ttgtagatgt cgttagctgg ggccacagcc
     3061 acaccgatga acagggcacc caacacgggc atgaagaact gcaagctatt ctcgctgcac
     3121 accacgatcc gatggtttgt attcagccca tagcgcttca tagcttctgc cagccgaacg
     3181 ctcatctcga agtactcggc gtaggtaatg tccacctcga tatgtgcgtc ggtaaaggcg
     3241 atggtgccgg gcaccagggc gtagcgcttc atggctttgt gcagctgctc gccggcggtc
     3301 ccgtcttcga gtgggtagaa tggcgctggg cccttcttaa tgtttttggc atcttccatg
     3361 cttaagcctg caggcgacct gcagttggac ctgggagtgg acacctgtgg agagaaaggc
     3421 aaagtggatg tcattgtcac tcaagtgtat ggccagatct caagcctgcc acacctcaag
     3481 cttgacaaca aaaagattgt cttttctgac cagatggacg cggccaccct caaaggcatc
     3541 accgcgggcc aggtgaatat caaatcctcc tcgtttttgg aaactgacaa tcttagcgca
     3601 gaagtcatgc ccgcttttga gagggagtac tcaccccaac agctggccct cgcagacagc
     3661 gaattaatcg cccgctcgct ccctctgcct ctcgctggaa ttactacagc gagttagata
     3721 aagccccgaa aaccggcttt tatactcagc gcgatccctc cctccgttct ttttcccgcc
     3781 aagcctctga gaagccctgc ccttctcgag gcaggagggg agccagggac ggccggggcc
     3841 cggcggtggc ggccgcgagc agcacagctc gggggtcctc agccgtccag accctaggga
     3901 gctcaggtac cggccagtta ggccagagaa atgttctggc acctgcactt gcactgggga
     3961 cagcctattt tgctagtttg ttttgtttcg ttttgttttg atggagagcg tatgttagta
     4021 ctatcgattc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     4081 ggcctgtcca atacctcccg taccttaata ttacttactt atccttgaga gacgtactag
     4141 taaccctgat aaatgcttca ataatattga aaaaggacga gtatgagcat ccaacatttt
     4201 cgtgtcgcac tcattccctt ctttgcggca ttttgcttgc ctgtttttgc acaccccgaa
     4261 acgctggtga aagtaaaaga tgctgaagat caactgggtg caagagtggg ctatatcgaa
     4321 ctggatctca atagcggcaa gatccttgag tctttccgcc ccgaagaacg attcccgatg
     4381 atgagcactt tcaaagtact gctatgtggc gcggtgttgt cccgtataga cgccgggcaa
     4441 gagcagcttg gtcgccgtat acactactca caaaacgact tggttgagta ctcgccggtc
     4501 acggaaaagc atcttacgga tggcatgacg gtaagagaat tgtgtagtgc tgccattacc
     4561 atgagcgaca acaccgcggc caacttactt ctgacaacga tcggaggccc taaggagctg
     4621 actgcatttc ttcataatat gggtgatcat gtgacccggc ttgaccgctg ggaaccagag
     4681 ttgaacgaag ccataccgaa cgacgagcgt gataccacga tgccagtagc aatggccaca
     4741 actcttcgga aactactcac tggcgaactt cttactctag catcacgaca gcagctcata
     4801 gactggatgg aggcggacaa agtagcagga ccacttcttc gctcggccct ccctgctggc
     4861 tggttcattg ctgacaaatc gggggccggt gaacgcggct ctcgcggcat cattgctgcg
     4921 ctggggcctg atggtaagcc ctcacgaatc gtagtgatct acacgacggg gagtcaggcc
     4981 actatggacg aacgaaatag gcagatcgct gagatcggtg cctcactgat caagcactgg
     5041 taaccactgc agtggtttag catttgcggc cgctgtcaga ccaagtttac tcatatatac
     5101 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5161 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5221 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5281 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5341 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5401 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5461 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5521 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5581 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5641 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5701 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5761 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5821 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5881 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5941 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6001 aggaagcgga agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     6061 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     6121 gctctcaagg gcatcggtcg gatccttatc gattttacca catttgtaga ggttttactt
     6181 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     6241 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6301 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6361 gtatcttatc atgtctgctc gaagcggccg gccgaattcg tcgaggatct agccattcta
     6421 gatcgaacaa gtttgtacaa aaaagctgaa cgagaa
//
