LOCUS       sequence-346540-        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 (R1-R2)..
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  JOURNAL   Unpublished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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..6321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1411..1430)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1413..1642
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     misc_feature    1798..1889
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(1830..1849)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    1903..1951
                     /note="synthetic polyadenylation signal"
     CDS             join(2056..2124,2125..2916)
                     /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      3116..3704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3605..3624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3858..3875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4120..4241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4157..4176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4211..4230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4300..4424
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4465..4770)
                     /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(4567..4586)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(5090..5770)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5685..5704
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5824..5830,5831..5848,5849..5854))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5879..6003)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(join(6036..6321,1..1367))
                     /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 ctctccagtt cggctggggc tacctggtag cccttgtatt tgatcaggct cttcagccgg
       61 tccacgatga agaagtgctc gtcctcgtcc cagtaggcga tgtcgccgct gtgcagccag
      121 ccgtccttgt cgatgagagc gtttgtagcc tcggggttgt taacgtagcc gctcatgatc
      181 atggggccac ggacgcacag ctcgccgcgc tggttcacac ccagtgtctt accggtgtcc
      241 aagtccacca ccttagcctc gaagaagggc accaccttgc ctactgcgcc aggcttgtcg
      301 tccccttcgg gggtgatcag aatggcgctg gttgtttctg tcaggccgta gccctggcgg
      361 atgcctggta ggtggaagcg tttggccacg gcctcaccta cctccttgct gagcggcgcc
      421 ccgccgctgg cgatctcgtg caagttgctt aggtcgtact tgtcgatgag agtgctctta
      481 gcgaagaagc taaatagtgt gggcaccagc agggcagatt gaatcttata gtcttgcaag
      541 ctgcgcaaga atagctcctc ctcgaagcgg tacatgagca cgacccgaaa gccgcagatc
      601 aagtagccca gcgtggtgaa catgccgaag ccgtggtgaa atggcaccac gctgaggata
      661 gcggtgtcgg ggatgatctg gttgccgaag atggggtcgc gggcatgact gaatcggaca
      721 caagcggtgc ggtgcggtag ggctacgccc ttgggcaatc cggtactgcc actactgttc
      781 atgatcaggg cgatggtttt gtcccggtcg aagctctcgg gcacgaagtc gtactcgttg
      841 aagccgggtg gcaaatggga agtcacgaag gtgtacatgc tttggaagcc ctggtagtcg
      901 gtcttgctat ccatgatgat gatcttttgt atgatcggta gcttcttttg cacgttgagg
      961 atcttttgca gccctttctt gctcacgaat acgacggtgg gctggctgat gcccatgctg
     1021 ttcagcagct cgcgctcgtt gtagatgtcg ttagctgggg ccacagccac accgatgaac
     1081 agggcaccca acacgggcat gaagaactgc aagctattct cgctgcacac cacgatccga
     1141 tggtttgtat tcagcccata gcgcttcata gcttctgcca gccgaacgct catctcgaag
     1201 tactcggcgt aggtaatgtc cacctcgata tgtgcgtcgg taaaggcgat ggtgccgggc
     1261 accagggcgt agcgcttcat ggctttgtgc agctgctcgc cggcggtccc gtcttcgagt
     1321 gggtagaatg gcgctgggcc cttcttaatg tttttggcat cttccatgct taagcctgca
     1381 ggcgacctgc agttggacct gggagtggac acctgtggag agaaaggcaa agtggatgtc
     1441 attgtcactc aagtgtatgg ccagatctca agcctgccac acctcaagct tgacaacaaa
     1501 aagattgtct tttctgacca gatggacgcg gccaccctca aaggcatcac cgcgggccag
     1561 gtgaatatca aatcctcctc gtttttggaa actgacaatc ttagcgcaga agtcatgccc
     1621 gcttttgaga gggagtactc accccaacag ctggccctcg cagacagcga attaattcgg
     1681 gccccggtcc gtaggcacgt ctgctcggct cgagtgttcg atcgcgactg aggacgaacg
     1741 cgcccccacc cccttatata agtaccctag ggagctcagg taccggccag ttaggccaga
     1801 gaaatgttct ggcacctgca cttgcactgg ggacagccta ttttgctagt ttgttttgtt
     1861 tcgttttgtt ttgatggaga gcgtatgtta gtactatcga ttcacacaaa aaaccaacac
     1921 acagatgtaa tgaaaataaa gatattttat tgcggcctgt ccaatacctc ccgtacctta
     1981 atattactta cttatccttg agagacgtac tagtaaccct gataaatgct tcaataatat
     2041 tgaaaaagga cgagtatgag catccaacat tttcgtgtcg cactcattcc cttctttgcg
     2101 gcattttgct tgcctgtttt tgcacacccc gaaacgctgg tgaaagtaaa agatgctgaa
     2161 gatcaactgg gtgcaagagt gggctatatc gaactggatc tcaatagcgg caagatcctt
     2221 gagtctttcc gccccgaaga acgattcccg atgatgagca ctttcaaagt actgctatgt
     2281 ggcgcggtgt tgtcccgtat agacgccggg caagagcagc ttggtcgccg tatacactac
     2341 tcacaaaacg acttggttga gtactcgccg gtcacggaaa agcatcttac ggatggcatg
     2401 acggtaagag aattgtgtag tgctgccatt accatgagcg acaacaccgc ggccaactta
     2461 cttctgacaa cgatcggagg ccctaaggag ctgactgcat ttcttcataa tatgggtgat
     2521 catgtgaccc ggcttgaccg ctgggaacca gagttgaacg aagccatacc gaacgacgag
     2581 cgtgatacca cgatgccagt agcaatggcc acaactcttc ggaaactact cactggcgaa
     2641 cttcttactc tagcatcacg acagcagctc atagactgga tggaggcgga caaagtagca
     2701 ggaccacttc ttcgctcggc cctccctgct ggctggttca ttgctgacaa atcgggggcc
     2761 ggtgaacgcg gctctcgcgg catcattgct gcgctggggc ctgatggtaa gccctcacga
     2821 atcgtagtga tctacacgac ggggagtcag gccactatgg acgaacgaaa taggcagatc
     2881 gctgagatcg gtgcctcact gatcaagcac tggtaaccac tgcagtggtt tagcatttgc
     2941 ggccgctgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3001 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3061 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3241 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc tgccggcacc
     3901 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     3961 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcggatcctt
     4021 atcgatttta ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4081 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg ctcgaagcgg
     4261 ccggccgaat tcgtcgacga tctagccatt ctagatcgaa ccactttgta caagaaagct
     4321 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa
     4381 cagactacat aatactgtaa aacacaacat atccagtcac tatggtcgac ctgcagactg
     4441 gctgtgtata agggagcctg acatttatat tccccagaac atcaggttaa tggcgttttt
     4501 gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg agaccggcac
     4561 actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca ccaccgggta
     4621 aagttcacgg gggactttat ctgacagcag acgtgcactg gccaggggga tcaccatccg
     4681 tcgcccgggc gtgtcaataa tatcactctg tacatccaca aacagacgat aacggctctc
     4741 tcttttatag gtgtaaacct taaactgcat ttcaccagcc cctgttctcg tcagcaaaag
     4801 agccgttcat ttcaataaac cgggcgacct cagccatccc ttcctgattt tccgctttcc
     4861 agcgttcggc acgcagacga cgggcttcat tctgcatggt tgtgcttacc agaccggaga
     4921 tattgacatc atatatgcct tgagcaactg atagctgtcg ctgtcaactg tcactgtaat
     4981 acgctgcttc atagcatacc tctttttgac atacttcggg tatacatatc agtatatatt
     5041 cttataccgc aaaaatcagc gcgcaaatac gcatactgtt atctggcttt tagtaagccg
     5101 gatcctctag attacgcccc gcctgccact catcgcagta ctgttgtaat tcattaagca
     5161 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     5221 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     5281 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     5341 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     5401 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     5461 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     5521 tcaccagctc accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg
     5581 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     5641 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     5701 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     5761 ttttctccat tttagcttcc ttagctcctg aaaatctcga cggatcctaa ctcaaaatcc
     5821 acacattata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gcggccgcca
     5881 tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta
     5941 atttaatata ttgatattta tatcatttta cgtttctcgt tcagcttttt tgtacaaact
     6001 tgtttgatca agctatcacc ggtgcatgct ctagattaca cggcgatctt gccgcccttc
     6061 ttggccttaa tgagaatctc gcggatcttg cgggcgtcca acttgccggt cagtccttta
     6121 ggcacctcgt ccacgaacac aacaccaccg cgcagcttct tggcggttgt aacctggctg
     6181 gccacatagt ccacgatctc cttctcggtc atggttttac cgtgttccag cacgacgact
     6241 gcggcgggca gctcgccggc atcgtcgtcg ggcaggccgg cgaccccggc gtcgaagatg
     6301 ttggggtgtt gcagcaggat g
//
