LOCUS       sequence-210554-        4263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recipient vector for expression of C-terminally Avi-tagged protein 
            of interest in chick embryos under the control of enhancer of 
            choice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4263)
  AUTHORS   Lukoseviciute M, Ling ITC, Senanayake U, Candido-Ferreira I, Taylor 
            G, Williams RM, Sauka-Spengler T
  TITLE     Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with 
            Sequencing in Zebrafish and Chicken.
  JOURNAL   STAR Protoc. 2020 Jul 31;1(2):100066. doi: 
            10.1016/j.xpro.2020.100066. eCollection 2020 Sep 18.
  PUBMED    33111104
REFERENCE   2  (bases 1 to 4263)
  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"; journalName: "STAR 
            Protoc."; date: "2020-07-31"; volume: "1(2):100066. doi"; pages: " 
            10.1016/j.xpro.2020.100066. eCollection 2020 Sep 18"
COMMENT     Sequence Label: pTK-ENH_BsmB1-MCS-Avi
FEATURES             Location/Qualifiers
     source          1..4263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(78..617)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLPGN
                     SSSVPIDREMFWHLHLHWGQPILLVCFVSFCFDGERMLVLSIHTKNQHTDVMKIKIFYC
                     GRSKYLPYLNITYLSW"
     polyA_signal    131..179
                     /note="synthetic polyadenylation signal"
     misc_feature    193..284
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     233..252
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     467..489
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     481..498
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     482..498
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     508..527
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        508..526
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(574..594)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(574..592)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(613..629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(613..629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(626..648)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    637..653
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(661..667,668..685,686..691))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    706..727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        895..1040
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1118..1250
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1233..1252
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1304..1313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1328..1351
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1400..1444
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     polyA_signal    1517..1638
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1528..1547)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1582..1601
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1886..1903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2057..2645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2137..2156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2816..3607,3608..3676))
                     /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     3439..3458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3677..3781)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3808..4236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3895..3914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4105..4126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg
      121 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
      181 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
      241 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag
      301 ctcgagttgc cagggagacg ggcgccgcta cagggcgcgt cccattcgcc attcaggctg
      361 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      421 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      481 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
      541 gggccccccc tcgaggtcct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
      601 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
      661 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
      721 cacattaatt gcgttgcgct cactgcccgc tttccaccgg tcgtctcctg ttggctagcc
      781 cgggctcgag atctgcgatc taagtaagct tgctgcttca tccccgtggc ccgttgctcg
      841 cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca
      901 caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg
      961 cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga
     1021 ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc agatcccaag cttgcggccg
     1081 cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
     1141 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
     1201 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
     1261 ccaggttcaa tacagctctt aagcggccgc aagctagctt gccgccacca tggtttacgt
     1321 aggaggtgac tacaaggacg acgacgacaa gggtggagca gctggaggtg agaacctgta
     1381 cttccagttc atggctggtg gcctgaatga catctttgag gcccagaaga tcgagtggca
     1441 tgaggactag taatctgtac aagtaagcgg ccgcgactct agagtcgggg cggccggccg
     1501 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1561 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1621 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1681 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     1741 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     1801 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     1861 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1921 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1981 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2041 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2101 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2161 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2221 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2281 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2341 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2401 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2461 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2521 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2581 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2641 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2701 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2761 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2821 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2881 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2941 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3001 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3061 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3121 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3181 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3241 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3301 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3361 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3421 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3481 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3541 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3601 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3661 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3781 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     3841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4261 ttt
//
