LOCUS       sequence-147009-        7660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian DamID vector for fusing gene of interest with Dam-V5 using
            Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7660)
  AUTHORS   Vogel MJ, Guelen L, de Wit E, Peric-Hupkes D, Loden M, Talhout W, 
            Feenstra M, Abbas B, Classen AK, van Steensel B
  TITLE     Human heterochromatin proteins form large domains containing 
            KRAB-ZNF genes.
  JOURNAL   Genome Res. 2006 Dec;16(12):1493-504. Epub 2006 Oct 12.
  PUBMED    17038565
REFERENCE   2  (bases 1 to 7660)
  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: "Genome 
            Res."; date: "2006-12"; volume: "16(12)"; pages: "1493-504. Epub 
            2006 Oct 12"
COMMENT     Sequence Label: pINDgw RFA-V5-EcoDam
FEATURES             Location/Qualifiers
     source          1..7660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        206..444
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     391..412
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    602..726
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(751..756,757..774,775..781)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             835..1494
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(901..920)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1836..2141
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2020..2039
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2182..2306)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2314..2355
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3208..3225
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3248..3265)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3254..3478
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3524..3952
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3611..3630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3821..3842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3961..3981)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3966..4296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4147..4282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4209..4228
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4363..5157
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4417..4436)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5027..5046
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5333..5454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5370..5389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5424..5443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5503..5519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5503..5519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5516..5538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5527..5543
                     /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(5551..5557,5558..5575,5576..5581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5596..5617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5734..5751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5905..6493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5985..6004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6664..7455,7456..7524))
                     /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     7287..7306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7525..7629)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctcgg ccgcatatta agtgcattgt tctcgatacc gctaagtgca
       61 ttgttctcgt tagctcgatg gacaagtgca ttgttctctt gctgaaagct cgatggacaa
      121 gtgcattgtt ctcttgctga aagctcgatg gacaagtgca ttgttctctt gctgaaagct
      181 cagtacccgg gagtaccctc gaccgccgga gtataaatag aggcgcttcg tctacggagc
      241 gacaattcaa ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa
      301 caagcgcagc tgaacaagct aaacaatctg cagtaaagtg caagttaaag tgaatcaatt
      361 aaaagtaacc agcaaccaag taaatcaact gcaactactg aaatctgcca agaagtaatt
      421 attgaataca agaagagaac tctgaatact ttcaacaagt taccgagaaa gaagaactca
      481 cacacagcta gcgtttaaac ttaagcttgg taccgagctc ggatccacta gtccagtgtg
      541 gtggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccttcgat
      601 cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
      661 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      721 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
      781 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
      841 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
      901 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
      961 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1021 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     1081 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1141 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1201 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1261 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1321 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1381 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1441 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     1501 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     1561 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     1621 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     1681 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     1741 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     1801 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     1861 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     1921 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     1981 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     2041 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     2101 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     2161 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gcattatgta
     2221 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2281 cgttcagctt tcttgtacaa agtggtgatc gaaggtaagc ctatccctaa ccctctcctc
     2341 ggtctcgatt ctacgcgtac cggcatgaag aaaaatcgcg cttttttgaa gtgggcaggg
     2401 ggcaagtatc ccctgcttga tgatattaaa cggcatttgc ccaagggcga atgtctggtt
     2461 gagccttttg taggtgccgg gtcggtgttt ctcaacaccg acttttctcg ttatatcctt
     2521 gccgatatca atagcgacct gatcagtctc tataacattg tgaagatgcg tactgatgag
     2581 tacgtacagg ccgcacgcga gctgtttgtt cccgaaacaa attgcgccga ggtttactat
     2641 cagttccgcg aagagttcaa caaaagccag gatccgttcc gtcgggcggt actgttttta
     2701 tatttgaacc gctacggtta caacggcctg tgtcgttaca atctgcgcgg tgagtttaac
     2761 gtgccgttcg gccgctacaa aaaaccctat ttcccggaag cagagttgta tcacttcgct
     2821 gaaaaagcgc agaatgcctt tttctattgt gagtcttacg ccgatagcat ggcgcgcgca
     2881 gatgatgcat ccgtcgtcta ttgcgatccg ccttatgcac cgctgtctgc gaccgccaac
     2941 tttacggcgt atcacacaaa cagttttacg cttgaacaac aagcgcatct ggcggagatc
     3001 gccgaaggtc tggttgagcg ccatattcca gtgctgatct ccaatcacga tacgatgtta
     3061 acgcgtgagt ggtatcagcg cgcaaaattg catgtcgtca aagttcgacg cagtataagc
     3121 agcaacggcg gcacacgtaa aaaggtggac gaactgctgg ctttgtacaa accaggagtc
     3181 gtttcacccg cgaaaaaata agccggtcat catcaccatc accattgagt ttaaacccgc
     3241 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3301 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3361 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3421 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3481 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3541 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3601 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3661 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3721 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3781 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3841 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     3901 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3961 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg
     4021 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4141 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4201 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4261 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4321 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4381 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4441 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4501 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4561 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4621 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4681 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4741 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4801 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4861 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4921 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4981 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5041 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5101 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5161 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5221 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5281 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5341 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5401 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5461 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5521 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5581 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5641 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5701 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5761 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5821 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5881 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5941 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6001 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6061 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6421 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6481 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6541 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6601 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6661 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6721 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6781 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6841 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6901 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6961 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7021 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7081 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7141 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7201 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7261 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7321 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7381 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7441 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7501 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7621 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
