LOCUS       sequence-194174-        5543 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression plasmid that allows for the expression of a 
            gene of interest fused to the N-terminus or C-terminus of ACP-tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5543)
  AUTHORS   Ana Egana
  TITLE     Egana Lab NEB plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5543)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pACP-tag (m)- 2 Vector
FEATURES             Location/Qualifiers
     source          1..5543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(156..173)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     383..402
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             598..1401
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(661..680)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1271..1290
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1531..1652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1568..1587)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1754..1770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1754..1770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1767..1789)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1778..1794
                     /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(1802..1808,1809..1826,1827..1832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1847..1868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1985..2002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2156..2744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2236..2255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2915..3706,3707..3775))
                     /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     3538..3557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3776..3880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3955..3974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4146..4525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4526..4729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4680..4700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4774..4793
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4774..4792
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4886..5119
                     /codon_start=1
                     /gene="acpP"
                     /product="acyl carrier protein (ACP) from E. coli"
                     /label=ACP
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     intron          5230..5459
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5442..5461
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(5538..5543,1..562)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
ORIGIN
        1 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
       61 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
      121 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
      181 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
      241 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
      301 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
      361 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
      421 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
      481 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
      541 cctttgaaaa acacgatgat aagcttgcca caacccggga taattcctgc agccaatatg
      601 ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
      661 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
      721 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
      781 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
      841 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
      901 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
      961 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     1021 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     1081 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     1141 cccgacggcg atgatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     1201 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     1261 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     1321 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     1381 cttcttgacg agttcttctg aggggatcaa ttctctagat aactgatcat aatcagccat
     1441 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1501 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1561 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1621 tgtggtttgt ccaaactcat caatgtatct taacgcgtcg agtgcattct agttgtggtt
     1681 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     1741 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1801 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1861 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2581 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2761 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2821 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2881 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2941 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3001 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3061 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3121 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3181 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3241 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3301 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3361 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3421 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3481 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3541 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3601 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3661 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3721 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3781 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3841 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3901 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     3961 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     4021 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     4081 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     4141 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     4201 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     4261 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     4321 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     4381 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     4441 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     4501 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     4561 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     4621 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     4681 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
     4741 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     4801 agcttggtac cgagctcgga tcgtcgctag cgatatcggc gcgccagcat ttaaatctgt
     4861 acagaccggt gaattcaagc ttaccatgag cactatcgaa gaacgcgtta agaaaattat
     4921 cggcgaacag ctgggcgtta agcaggaaga agttaccaac aatgcttctt tcgttgaaga
     4981 cctgggcgcg gattctcttg acaccgttga gctggtaatg gctctggaag aagagtttga
     5041 tactgagatt ccggacgaag aagctgagaa aatcaccacc gttcaggctg ccattgatta
     5101 catcaacggc caccaggcgc ctgcaggcgg atccgcgttt aaactcgagg ttaattaatg
     5161 agcggccgca tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca
     5221 gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag
     5281 tttccaaaaa cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg
     5341 ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc
     5401 ttgagatctg gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg
     5461 tgtccactcc caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc
     5521 tctccccccc ccccttttcc ctc
//
