LOCUS       sequence-346549-        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 (R1-R2)..
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  JOURNAL   Unpublished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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             187..492
                     /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     371..390
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(533..657)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    716..837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(727..746)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     781..800
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1082..1099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1253..1841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1333..1352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2041..2832,2833..2901))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3006..3054
                     /note="synthetic polyadenylation signal"
     misc_feature    3068..3159
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3108..3127
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     intron          3450..3679
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3662..3681
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3725..5377
                     /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"
     protein_bind    5410..5534
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5559..5564,5565..5582,5583..5589)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5643..6323
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(5709..5728)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
       61 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
      121 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
      181 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
      241 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
      301 gcacgtctgc tgtcagataa agtcccccgt gaactttacc cggtggtgca tatcggggat
      361 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
      421 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
      481 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
      541 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
      601 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggttcg
      661 atctagaatg gctagatcgt cgacgaattc ggccggccgc ttcgagcaga catgataaga
      721 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
      781 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
      841 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
      901 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgaccga tgcccttgag
      961 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     1021 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     1081 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1141 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1201 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1261 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1321 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1381 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1441 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1501 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1561 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1621 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1681 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1741 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1801 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1861 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1921 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1981 aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg
     2041 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
     2101 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
     2161 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
     2221 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
     2281 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
     2341 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
     2401 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
     2461 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
     2521 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
     2581 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
     2641 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
     2701 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
     2761 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
     2821 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
     2881 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
     2941 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
     3001 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     3061 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     3121 ctgtccccag tgcaagtgca ggtgccagaa catttctctg gcctaactgg ccggtacctg
     3181 agctccctag ggtctggacg gctgaggacc cccgagctgt gctgctcgcg gccgccaccg
     3241 ccgggccccg gccgtccctg gctcccctcc tgcctcgaga agggcagggc ttctcagagg
     3301 cttggcggga aaaagaacgg agggagggat cgcgctgagt ataaaagccg gttttcgggg
     3361 ctttatctaa ctcgctgtag taattccagc gagaggcaga gggagcgagc gggcgattaa
     3421 ttcgctgtct gcgagggcca gctgttgggg tgagtactcc ctctcaaaag cgggcatgac
     3481 ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc
     3541 ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc
     3601 aagcttgagg tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac
     3661 tttgcctttc tctccacagg tgtccactcc caggtccaac tgcaggtcgc ctgcaggctt
     3721 aagcatggaa gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga
     3781 cgggaccgcc ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac
     3841 catcgccttt accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat
     3901 gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt
     3961 ggtgtgcagc gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg
     4021 tgtggctgtg gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg
     4081 catcagccag cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt
     4141 gcaaaagaag ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca
     4201 gggcttccaa agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta
     4261 cgacttcgtg cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag
     4321 tggcagtacc ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt
     4381 cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag
     4441 cgtggtgcca tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg
     4501 ctttcgggtc gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga
     4561 ctataagatt caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac
     4621 tctcatcgac aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct
     4681 cagcaaggag gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg
     4741 ctacggcctg acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc
     4801 tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg
     4861 taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag
     4921 cggctacgtt aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca
     4981 cagcggcgac atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa
     5041 gagcctgatc aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct
     5101 gcaacacccc aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga
     5161 gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt
     5221 ggactatgtg gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt
     5281 ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct
     5341 cattaaggcc aagaagggcg gcaagatcgc cgtgtaatct agagcatgca ccggtgatag
     5401 cttgatcaaa caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     5461 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     5521 atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct
     5581 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
     5641 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     5701 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     5761 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     5821 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     5881 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     5941 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     6001 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     6061 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     6121 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     6181 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     6241 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag gcggggcgta
     6301 atctagagga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     6361 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct
     6421 atgaagcagc gtattacagt gacagttgac agcgac
//
