LOCUS       sequence-346541-        6321 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 an SCP1 
            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 6321)
  TITLE     Transcription imparts architecture, function, and logic to enhancer 
            units
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6321)
  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            "Unpublished"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-ccw
FEATURES             Location/Qualifiers
     source          1..6321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    699..823
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(864..1169)
                     /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     complement(966..985)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1489..2169)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     2084..2103
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2223..2229,2230..2247,2248..2253))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2278..2402)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2461..2582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2472..2491)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2526..2545
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2827..2844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2998..3586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3078..3097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3786..4577,4578..4646))
                     /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    4751..4799
                     /note="synthetic polyadenylation signal"
     misc_feature    4813..4904
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4853..4872
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     intron          5060..5289
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5272..5291
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             join(5335..6321,1..666)
                     /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"
ORIGIN
        1 cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac cagcgccatt
       61 ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc
      121 gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca gcgcggcgag
      181 ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga ggctacaaac
      241 gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg ggacgaggac
      301 gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg ctaccaggta
      361 gccccagccg aactggagag catcctgctg caacacccca acatcttcga cgccggggtc
      421 gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac
      481 ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc
      541 aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag
      601 ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg caagatcgcc
      661 gtgtaatcta gagcatgcac cggtgatagc ttgatcaaac cactttgtac aagaaagctg
      721 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
      781 agactacata atactgtaaa acacaacata tccagtcact atggtcgacc tgcagactgg
      841 ctgtgtataa gggagcctga catttatatt ccccagaaca tcaggttaat ggcgtttttg
      901 atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga gaccggcaca
      961 ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac caccgggtaa
     1021 agttcacggg ggactttatc tgacagcaga cgtgcactgg ccagggggat caccatccgt
     1081 cgcccgggcg tgtcaataat atcactctgt acatccacaa acagacgata acggctctct
     1141 cttttatagg tgtaaacctt aaactgcatt tcaccagccc ctgttctcgt cagcaaaaga
     1201 gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt ccgctttcca
     1261 gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca gaccggagat
     1321 attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt cactgtaata
     1381 cgctgcttca tagcatacct ctttttgaca tacttcgggt atacatatca gtatatattc
     1441 ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggctttt agtaagccgg
     1501 atcctctaga ttacgccccg cctgccactc atcgcagtac tgttgtaatt cattaagcat
     1561 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     1621 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     1681 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     1741 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     1801 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     1861 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     1921 caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc
     1981 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     2041 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     2101 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     2161 tttctccatt ttagcttcct tagctcctga aaatctcgac ggatcctaac tcaaaatcca
     2221 cacattatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg cggccgccat
     2281 agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa
     2341 tttaatatat tgatatttat atcattttac gtttctcgtt cagctttttt gtacaaactt
     2401 gttcgatcta gaatggctag atcctcgacg aattcggccg gccgcttcga gcagacatga
     2461 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2521 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2581 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2641 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg accgatgccc
     2701 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     2761 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     2821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3421 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3721 aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct aaaccactgc
     3781 agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct atttcgttcg
     3841 tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg cttaccatca
     3901 ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga tttgtcagca
     3961 atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt gtccgcctcc
     4021 atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt gagtagtttc
     4081 cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt cggtatggct
     4141 tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat attatgaaga
     4201 aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg
     4261 tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc cgtaagatgc
     4321 ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat acggcgacca
     4381 agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag tactttgaaa
     4441 gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt gccgctattg
     4501 agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc ttttactttc
     4561 accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt
     4621 gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg aagcatttat
     4681 cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg ggaggtattg
     4741 gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     4801 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
     4861 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctggccta actggccggt
     4921 acctgagctc cctagggtac ttatataagg gggtgggggc gcgttcgtcc tcagtcgcga
     4981 tcgaacactc gagccgagca gacgtgccta cggaccgggg cccgaattaa ttcgctgtct
     5041 gcgagggcca gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta
     5101 agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc ggtgatgcct
     5161 ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc aagcttgagg
     5221 tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac tttgcctttc
     5281 tctccacagg tgtccactcc caggtccaac tgcaggtcgc ctgcaggctt aagcatggaa
     5341 gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc
     5401 ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt
     5461 accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg
     5521 ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc
     5581 gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg
     5641 gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag
     5701 cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag
     5761 ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa
     5821 agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg
     5881 cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc
     5941 ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc
     6001 cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca
     6061 tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc
     6121 gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt
     6181 caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac
     6241 aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag
     6301 gtaggtgagg ccgtggccaa a
//
