LOCUS       sequence-166523-        6292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAR domain of human Amphiphysin1 (amino acids 1-256), C-terminally 
            tagged with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6292)
  AUTHORS   Hayer A, Shao L, Chung M, Joubert LM, Yang HW, Tsai FC, Bisaria A, 
            Betzig E, Meyer T
  TITLE     Engulfed cadherin fingers are polarized junctional structures 
            between collectively migrating endothelial cells.
  JOURNAL   Nat Cell Biol. 2016 Dec;18(12):1311-1323. doi: 10.1038/ncb3438. Epub
            2016 Nov 14.
  PUBMED    27842057
REFERENCE   2  (bases 1 to 6292)
  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: "10.1038/ncb3438"; 
            journalName: "Nat Cell Biol"; date: "2016-12"; volume: "18"; issue: 
            "12"; pages: "1311-1323"
COMMENT     Sequence Label: pCAG-AMPH1-BAR-mCherry
FEATURES             Location/Qualifiers
     source          1..6292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2502..2511
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2508..3218
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2653..2671)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2921..2940
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3316..3335)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3381..3436
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3435..3454)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3795..3811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3795..3811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3808..3830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3819..3835
                     /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(3843..3849,3850..3867,3868..3873))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3888..3909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3968..4163
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4014..4149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4169..4303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4206..4225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4260..4279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4371..4388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4542..5130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4622..4641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5301..6092,6093..6161))
                     /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     5924..5943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6162..6266)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcatggcc gacatcaaga
     1741 cgggcatctt cgccaagaac gtccagaagc gactcaaccg cgcgcaggaa aaggtcctcc
     1801 aaaagctggg gaaagctgat gagacaaaag acgaacagtt cgaagaatat gtccagaact
     1861 tcaaacggca agaagcagag ggtaccagac ttcagcgaga actccgagga tatttagcag
     1921 caatcaaagg catgcaggag gcctccatga agctcacaga gtcgctgcat gaagtctatg
     1981 agcctgactg gtatgggcgg gaagatgtga aaatggttgg tgagaaatgt gatgtgctgt
     2041 gggaagactt ccatcaaaaa ctcgtggatg ggtccttgct aacactggat acctacctgg
     2101 ggcaatttcc tgacataaag aatcgcatcg ccaagcgcag caggaagcta gtggactatg
     2161 acagtgcccg ccaccatctg gaagctctgc agagctccaa gaggaaggat gagagtcgaa
     2221 tctctaaggc agaagaagaa tttcagaaag cacagaaagt gtttgaagag tttaacgttg
     2281 acttacaaga agagttacca tcattatggt caagacgagt tggattttat gttaatactt
     2341 tcaaaaacgt ctccagcctt gaagccaagt ttcataagga aattgcggtg ctttgccaca
     2401 aactgtatga agtgatgaca aaactgggtg accagcacgc cgacaaggcc ttcaccatcc
     2461 aaggagcgcc cagtgattcg ggtcctctcc gcggaccggt cgccaccatg gtgagcaagg
     2521 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     2581 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2641 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2701 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     2761 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2821 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     2881 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2941 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     3001 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     3061 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     3121 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     3181 gccactccac cggcggcatg gacgagctgt acaagtaact cgaggatatc gcggccgcac
     3241 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     3301 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3361 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3421 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     3481 tgagtatttg gtttagagtt tggcaacata tgccatatgc tggctgccat gaacaaaggt
     3541 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     3601 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     3661 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     3721 tactcccagt catagctgtc cctcttctct tatgaagatc cctcgacctg cagcccaagc
     3781 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3841 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3901 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3961 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4021 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4081 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4141 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
     4201 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4261 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
     4321 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4381 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4441 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4501 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4561 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4621 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4681 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     4741 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4801 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4861 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4921 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4981 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5041 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5101 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5161 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5221 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5281 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5341 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5401 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5461 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5521 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5581 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5641 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5701 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5761 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5821 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5881 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5941 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6001 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6061 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6121 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6181 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6241 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tg
//
