LOCUS       sequence-258507-        5505 bp    DNA     circular SYN 15-APR-2025
DEFINITION  enhancer reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5505)
  AUTHORS   Williams RM, Candido-Ferreira I, Repapi E, Gavriouchkina D, 
            Senanayake U, Ling ITC, Telenius J, Taylor S, Hughes J, 
            Sauka-Spengler T
  TITLE     Reconstruction of the Global Neural Crest Gene Regulatory Network In
            Vivo.
  JOURNAL   Dev Cell. 2019 Oct 21;51(2):255-276.e7. doi: 
            10.1016/j.devcel.2019.10.003.
  PUBMED    31639368
REFERENCE   2  (bases 1 to 5505)
  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"; doi: 
            "10.1016/j.devcel.2019.10"; journalName: "Dev Cell"; date: 
            "2019-10-21- 21"; volume: "51"; issue: "2"; pages: "255-276.e7"
COMMENT     Sequence Label: pTK_Pax7_enh_216_Cerulean
FEATURES             Location/Qualifiers
     source          1..5505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..105
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     54..73
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1276..1421
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1499..1631
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1614..1633
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1685..1694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1691..1693,1694..1696,1697..2410)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1736..1757)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1997..2016)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2344..2365
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2580..2701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2591..2610)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2645..2664
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2949..2966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3120..3708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3200..3219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3879..4670,4671..4739))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4502..4521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4740..4844)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4871..5326
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4958..4977)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5168..5189
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5457..5505
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttc catggcggtg tctggcatgg aaagagcagc tgctcggaga gagaataagt
      181 attagaagaa agagaaatcc tcgtaatctt gggacgtata taatctacgt cgccccgaat
      241 cccaaaaggc agcgggcccg tctctctgat ttatttttat gctagcctag tttgcatccc
      301 aaagaagtaa actgctattt ttaaccactg gtttaaagct taattaaaag cccctaaact
      361 gattgttttg aaaattgatt tagaggaatt gccccagggc tgagacgcta tctaagccaa
      421 gttgcagtcc cagagctcgc agtggtgtct gaatcgttaa ccccgcagag ccgggagtcg
      481 ccggggctgg atgacacatg ggctggccga gctccttgca ccctctgcaa agaaaggtcc
      541 tttgttaggc caggctactg gtttgctcgg gtggatctcc gctggtttcc tggcgttccg
      601 aggtttaatt gaattaaggc agagctcgga gatgctggtg acaccgggac ggagagctct
      661 gcctttcatc tgccggaggg gaatgtggct gggtcggggg acgtggaatt tggggacggg
      721 acctcgccat tcgatcgaag ggagctttac ctgctgaaaa ggagatggaa gttggttgaa
      781 ggcagggggg ctgcggaagg gaagtggcat ttcggcccct taattagatg taaacaagaa
      841 tagggacggg gctccatggt cagacaactt gattgttaag ggattaacac ctaagtaaac
      901 caataaaggc tctgcggggc tggtgagagg gctgcgggcc aggcgagccc ggcacactaa
      961 tgggttcccc ccggcacgac ccaccccaaa acggccctct ccgccccgcc agcgactttt
     1021 ggagcccggt tttaaggcta taagccaagg cttgagttat ttcttgctgg ccgtgctcct
     1081 tggaaagccc ccttcgggat gcttcaaggt gaggagcctc tcattaccgg cacccatcgc
     1141 cctaattaag gaccgctagc ccgggctcga gatctgcgat ctaagtaagc ttgctgcttc
     1201 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     1261 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     1321 acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt
     1381 gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg
     1441 cagatcccaa gcttgcggcc gcttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     1501 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     1561 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     1621 tctctccaca ggtgtccact cccaggttca atacagctct taagcggccg caagctagct
     1681 tgccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1741 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1801 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1861 ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc gctaccccga
     1921 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1981 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2041 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2101 cctggggcac aagctggagt acaacgccat cagcgacaac gtctatatca ccgccgacaa
     2161 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt
     2221 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2281 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     2341 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2401 gtacaagtga gcctctagac tccctcagcg gcgcgccacg ttggctgtag agtcgccagg
     2461 gcgtatttaa ctggagggac tctcggttag agttgatacg gtcccgtcgc atacagcgcc
     2521 tttggccaag atctcacgtg tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     2581 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2641 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2701 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2761 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     2821 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     2881 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2941 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3001 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3061 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3121 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3181 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3241 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3301 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3361 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3421 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3481 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3541 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3601 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3661 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3721 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3781 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3841 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3901 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3961 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4021 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4081 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4141 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4201 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4261 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4321 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4381 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4441 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4501 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4561 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4621 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4681 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4741 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4801 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4861 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4921 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4981 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5041 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5101 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5161 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5221 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5281 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc
     5341 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc
     5401 aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata
     5461 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtg
//
