LOCUS       sequence-249980-        5075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Preprotachykinin A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5075)
  TITLE     Michael Kressel plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5075)
  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            "Unpublished"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: Preprotachykinin A
FEATURES             Location/Qualifiers
     source          1..5075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1037..1046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1043..1741
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     polyA_signal    1863..1984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1900..1919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1954..1973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1991..2446)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2128..2149)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2340..2359
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2473..2577
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2579..2936
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2579..2770
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2602..2622)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2787..2922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2971..3765
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2971..3765
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3025..3044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3635..3654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3953..3972)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3997..4044
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4373..4961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4862..4881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcatgaaa atcctcgtgg ccttggcagt
      661 cttttttctt gtctccactc agctgtttgc agaagaaata ggagccaatg atgatctgaa
      721 ttactggtcc gactggtacg acagcgacca gatcaaggag gaactgccgg agccctttga
      781 gcatcttctg cagagaatcg cccggagacc caagcctcag cagttctttg gattaatggg
      841 caaacgggat gctgattcct caattgaaaa acaagtggcc ctgttaaagg ctctttatgg
      901 acatggccag atctctcaca aaagacataa aacagattcc tttgttggac taatgggcaa
      961 aagagcttta aattctgtgg cttatgaaag gagtgcaatg cagaattatg aaagaagacg
     1021 tggggatcca ccggtcgcca ccatggtgag cgagctgatt aaggagaaca tgcccatgaa
     1081 gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg
     1141 caagccctac gagggcaccc agaccatgag aatcaaggtc gtcgagggcg gccctctccc
     1201 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcagaacct tcatcaagca
     1261 ccctccgggc atccccgact tctttaagca gtccttccct gagggcttca catgggagag
     1321 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     1381 cggctgcctc atctacaacg tcaaggttag aggggtgaac ttcccagcca acggccctgt
     1441 gatgcagaag aaaacactcg gctgggaggc ctccaccgag acgatgtacc ccgctgacgg
     1501 cggcctggaa ggcgcatgtg acatggccct gaagctcgtg ggcgggggcc acctgatctg
     1561 caaccttgag accacataca gatccaagaa acccgctacg aacctcaaga tgcccggcgt
     1621 ctacaacgtg gaccacagac tggaaagaat caaggaggcc gacgatgaga cctacgtcga
     1681 gcagcacgag gtggctgtgg ccagatactc tactggtggc gctggtgatg gaggtaaatg
     1741 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1801 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1861 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1921 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1981 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2041 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2101 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2161 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2221 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2281 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2341 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2401 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2461 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2521 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2581 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2641 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2761 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2821 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     2941 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3001 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3061 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3121 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3181 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3241 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3301 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3361 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3421 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3481 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3541 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3601 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3661 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3721 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3781 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3841 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3901 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     3961 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4021 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4081 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4141 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4201 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4261 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4321 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4381 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4441 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4501 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4561 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4621 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4681 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4741 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4801 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4861 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4921 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4981 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5041 ctgattctgt ggataaccgt attaccgcca tgcat
//
