LOCUS       sequence-258451-        5546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enhancer reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5546)
  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 5546)
  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_TFAP2b_enh_226_Citrine
FEATURES             Location/Qualifiers
     source          1..5546
                     /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        1316..1461
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1539..1671
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1654..1673
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1725..1734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1731..1733,1734..1736,1737..2447)
                     /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(1776..1797)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2037..2056)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2384..2405
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2621..2742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2632..2651)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2686..2705
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2990..3007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3161..3749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3241..3260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3920..4711,4712..4780))
                     /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     4543..4562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4781..4885)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4912..5367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4999..5018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5209..5230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5498..5546
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggctggc tgtagatact ccgcgtgtat gtgtgtgtat acatacatgt
      181 actttggtta ctgacctgcg ggaaatattt tacagctact tatctctccc ggcgtagaga
      241 atgaaggcga caggacaggg tcgtttcctc ctttctgcct ctttctccac tccccgctca
      301 cactcagcag ccctatcccg tgcaggcagc atttcgccgt ccccttctat ttacccccaa
      361 cagctgaagg gctccttctg caatctgagc cgcgactgct gccagagggg gtgaccacgg
      421 tgggacaggg agctggaagg ggaagtaagg ctggggtata tctatagaaa tcccttctct
      481 atatctatag atagatatac atatacacgc atatatacat atatatgcat acggatatat
      541 ataaactccc tgctaaagtt gggaggggaa gatctgagta ggagagcatc cctccccctg
      601 ccccgtcccc tcggagcggg ctgataacga gcagcccctg cccgctcccc tttcccccct
      661 ccttcccctc acccccatcc caacatcctg tcaggcaata ggctgtaaaa tagttatcca
      721 gcccagcaca caggcaagtg ccgtttgaga gtaatttctg ccccgagcgt ttggccgggg
      781 gcaaagcaag cgtgctctct aactgatcaa cctggatatt tcattattca ttaatttact
      841 gtttaaccaa atcccactca ttatcagagg caacgcttgg cagagcccag cgaacagtag
      901 cagttggtgt caggcgtgga gaattggccc atttgatcct gggctgtttt attattcaac
      961 accactgact gagaaaaagg cccatgaata ataaagagcc aaagcgtgtt tgaactgaga
     1021 agcatgacac agccttacaa aacagcagtt aggaaattaa aggcgaggtg gcccaggctt
     1081 tgcatttttt attctcactg tgggcacggg cactgggatc ccatattttt tttgtccctc
     1141 ccgtttatcg ttgctgttat taccagtttc cccataagcc cttcgagact gttggctagc
     1201 ccgggctcga gatctgcgat ctaagtaagc ttgctgcttc atccccgtgg cccgttgctc
     1261 gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac
     1321 acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg
     1381 gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg
     1441 accctgcagc gacccgctta acagcgtcaa cagcgtgccg cagatcccaa gcttgcggcc
     1501 gcttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1561 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1621 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1681 cccaggttca atacagctct taagcggccg caagctagct tgccgccacc atggtgagca
     1741 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1801 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1861 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1921 ccttcggcta cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1981 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2041 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2101 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2161 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2221 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2281 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     2341 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2401 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga gcctctagac
     2461 tccctcagcg gcgcgccgtt gcgttccaaa cgtcgtggag ctccctgtca agacacgaac
     2521 cgctaacgcc accttagtcc gtgaagcatg atacgcgaca taccccagga gatctcacgt
     2581 gtctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     2641 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2701 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2761 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     2821 tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc
     2881 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2941 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     3001 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3061 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3121 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3601 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3661 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3721 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3781 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3841 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3901 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3961 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4021 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4081 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4141 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4201 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4261 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4321 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4381 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4441 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4501 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4561 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4621 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4681 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4741 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4801 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4861 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     4921 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4981 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5041 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5101 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5161 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5221 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5281 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5341 ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg
     5401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
     5461 gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt
     5521 acatctgtgt gttggttttt tgtgtg
//
