LOCUS       sequence-423104-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for transient expression of NAT1 gene with C-terminal HA tag,
            gene flanked by att sites for gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  TITLE     Chromosome 8p-engineering reveals increased metastasis targetable by
            patient-specific synthetic lethality
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6692)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDEST26-NAT1-C-HA
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     473..493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     519..543
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    640..664
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1535..1559)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1564..1590
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1621..1640)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1622..1640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1651..1668)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1651..1667)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1660..1682)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2072..2206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2109..2128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2163..2182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2332..2787
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2629..2650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2904..3261
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2904..3095
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2927..2947)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3112..3247
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3312..4106
                     /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(3366..3385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3976..3995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4170..4218
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4259..4278)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4378..4400
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4524..4628
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4629..4697,4698..5489)
                     /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     complement(4847..4866)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5660..6248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6149..6168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6402..6419
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6493..6597)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      601 agcctccgga ctctagccta ggccgcggag ctcgagtcaa caagtttgta caaaaaagtt
      661 ggcatggaca ttgaagcata tcttgaaaga attggctata agaagtctag gaacaaattg
      721 gacttggaaa cattaactga cattcttcaa caccagatcc gagctgttcc ctttgagaac
      781 cttaacatcc attgtgggga tgccatggac ttaggcttag aggccatttt tgatcaagtt
      841 gtgagaagaa atcggggtgg atggtgtctc caggtcaatc atcttctgta ctgggctctg
      901 accactattg gttttgagac cacgatgttg ggagggtatg tttacagcac tccagccaaa
      961 aaatacagca ctggcatgat tcaccttctc ctgcaggtga ccattgatgg caggaactac
     1021 attgtcgatg ctgggtttgg acgctcatac cagatgtggc agcctctgga gttaatttct
     1081 gggaaggatc agcctcaggt gccttgtgtc ttccgtttga cggaagagaa tggattctgg
     1141 tatctagacc aaatcagaag ggaacagtac attccaaatg aagaatttct tcattctgat
     1201 ctcctagaag acagcaaata ccgaaaaatc tactccttta ctcttaagcc tcgaacaatt
     1261 gaagattttg agtctatgaa tacatacctg cagacatctc catcatctgt gtttactagt
     1321 aaatcatttt gttccttgca gaccccagat ggggttcact gtttggtggg cttcaccctc
     1381 acccatagga gattcaatta taaggacaat acagatctaa tagagttcaa gactctgagt
     1441 gaggaagaaa tagaaaaagt gctgaaaaat atatttaata tttccttgca gagaaagctt
     1501 gtgcccaaac atggtgatag attttttact atttacccaa ctttcttgta caaagtggtt
     1561 gattacccat acgatgttcc agattacgct tgacgcgtgc atgcgacgtc atagctctct
     1621 ccctatagtg agtcgtatta taagctaggc actggccgtc gttttacaac gtcgtgactg
     1681 ggaaaactgc tagcttggga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
     1741 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
     1801 atgtgttaaa ctagctgcat atgcttgctg cttgagagtt ttgcttactg agtatgattt
     1861 atgaaaatat tatacacagg agctagtgat tctaattgtt tgtgtatttt agattcacag
     1921 tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca
     1981 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2041 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2101 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2161 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcgatc ctgcattaat
     2221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg ctggcgtaat agcgaagagg
     2281 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2341 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2401 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2461 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2521 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2581 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2641 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2701 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2761 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     2821 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     2881 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     2941 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3001 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3061 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3121 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3181 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3241 ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagg
     3301 ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3361 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3421 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3481 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3541 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ttggctgcta ttgggcgaag
     3601 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3661 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3721 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3781 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3841 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3901 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3961 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4021 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4081 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4141 gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     4201 gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc actctcagta
     4261 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4321 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4381 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4441 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4501 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4561 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4621 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4681 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4741 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4801 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4861 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4921 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4981 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5041 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5101 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5161 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5221 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5281 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5341 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5401 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5461 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5521 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5581 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5641 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5701 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5761 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5821 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5881 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5941 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6001 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6061 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6121 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6181 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6241 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6301 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6361 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6421 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6481 gcagagcttg caattcgcgc tttttcaata ttattgaagc atttatcagg gttattgtct
     6541 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6601 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6661 taaaaatagg cgtattacga ggccctttca cg
//
