LOCUS       sequence-249877-        4526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer cloning nanotag reporter vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4526)
  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 4526)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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_Citrine_Nanotag_19_neg_Control
FEATURES             Location/Qualifiers
     source          1..4526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        346..491
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          569..701
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     684..703
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      755..764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(761..763,764..766,767..1477)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(806..827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1067..1086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1414..1435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1650..1771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1661..1680)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1715..1734
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2019..2036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2190..2778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2270..2289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2949..3740,3741..3809))
                     /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     3572..3591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3810..3914)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3941..4396
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4028..4047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4238..4259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcgagttgc
      181 caggagctgg atcgatgata tccgatcgat cgtagcacct gttggctagc ccgggctcga
      241 gatctgcgat ctaagtaagc ttgctgcttc atccccgtgg cccgttgctc gcgtttgctg
      301 gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg
      361 cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc
      421 aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc
      481 gacccgctta acagcgtcaa cagcgtgccg cagatcccaa gcttgcggcc gcttaagctg
      541 cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg
      601 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
      661 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact cccaggttca
      721 atacagctct taagcggccg caagctagct tgccgccacc atggtgagca agggcgagga
      781 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      841 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      901 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
      961 cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1021 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1081 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1141 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1201 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1261 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1321 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     1381 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1441 cgccgggatc actctcggca tggacgagct gtacaagtga gcctctagac tccctcagcg
     1501 gcgcgcccaa tgggtgtatt gtgggttgcc gatttacggc gagtcttccg acggatatgg
     1561 acattgccgg aacagcctgt actggagcga ccttacggag atctcacgtg tctagagtcg
     1621 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     1681 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1741 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1801 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1861 taaaatcgat aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     1921 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     1981 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     2041 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2101 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2281 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2341 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     2401 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2461 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2521 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2581 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2641 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2701 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2761 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2821 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2881 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2941 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3001 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3061 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3121 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3181 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3241 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3301 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3361 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3421 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3481 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3541 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3601 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3661 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3721 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3781 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3841 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3901 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     3961 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4021 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4081 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4141 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4201 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4261 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4321 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4381 ttaacgctta caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4441 gtgcgggcct cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag
     4501 gtacgggagg tacttggagc ggccgc
//
