LOCUS       sequence-200644-        6413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of solute carrier family 25 member 27 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6413)
  AUTHORS   Shu Leong Ho
  TITLE     Shu Leong Ho Plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6413)
  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            "unpublished"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: pcDNA3.1-hUCP4-NE
FEATURES             Location/Qualifiers
     source          1..6413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1770..1787)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1776..2000
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2046..2474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2133..2152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2343..2364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2483..2503)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2488..2817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2668..2803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2730..2749
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2884..3678
                     /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(2938..2957)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3548..3567
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3852..3973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3889..3908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3943..3962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4022..4038)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4022..4038)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4035..4057)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4046..4062
                     /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(4070..4076,4077..4094,4095..4100))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4115..4136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4253..4270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4424..5012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4504..4523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5183..5974,5975..6043))
                     /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     5806..5825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6044..6148)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6223..6242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc gccaccatgt ccgtcccgga
      721 ggaggaggag aggcttttgc cgctgaccca gagatggccc cgagcgagca aattcctact
      781 gtccggctgc gcggctaccg tggccgagct agcaaccttt cccctggatc tcacaaaaac
      841 tcgactccaa atgcaaggag aagcagctct tgctcggttg ggagacggtg caagagaatc
      901 tgccccctat aggggaatgg tgcgcacagc cctagggatc attgaagagg aaggctttct
      961 aaagctttgg caaggagtga cacccgccat ttacagacac gtagtgtatt ctggaggtcg
     1021 aatggtcaca tatgaacatc tccgagaggt tgtgtttggc aaaagtgaag atgagcatta
     1081 tcccctttgg aaatcagtca ttggagggat gatggctggt gttattggcc agtttttagc
     1141 caatccaact gacctagtga aggttcagat gcaaatggaa ggaaaaagga aactggaagg
     1201 aaaaccattg cgatttcgtg gtgtacatca tgcatttgca aaaatcttag ctgaaggagg
     1261 aatacgaggg ctttgggcag gctgggtacc caatatacaa agagcagcac tggtgaatat
     1321 gggagattta accacttatg atacagtgaa acactacttg gtattgaata caccacttga
     1381 ggacaatatc atgactcacg gtttatcaag tttatgttct ggactggtag cttctattct
     1441 gggaacacca gccgatgtca tcaaaagcag aataatgaat caaccacgag ataaacaagg
     1501 aaggggactt ttgtataaat catcgactga ctgcttgatt caggctgttc aaggtgaagg
     1561 attcatgagt ctatataaag gctttttacc atcttggctg agaatgaccc cttggtcaat
     1621 ggtgttctgg cttacttatg aaaaaatcag agagatgagt ggagtcagtc catttctcga
     1681 gtctagaacc aaagaaaacc cgcgtagcaa ccaggaagaa agctatgatg ataacgaaag
     1741 ctaagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     1801 tattgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1861 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1921 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1981 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2041 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2101 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2161 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2221 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2281 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2341 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2401 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2461 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2521 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2581 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2641 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2701 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     2761 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     2821 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     2881 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2941 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3001 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3061 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3121 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3181 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3241 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3301 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3361 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3421 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3481 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3541 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3601 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3661 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3721 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3781 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3841 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3901 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3961 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4021 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4081 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4141 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4201 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4261 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4321 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4381 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4441 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4501 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4561 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4621 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4681 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4741 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4801 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4861 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4921 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4981 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5041 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5101 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5161 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5221 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5281 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5341 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5401 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5461 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5521 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5581 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5641 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5701 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5761 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5821 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5881 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5941 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6001 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6061 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6121 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     6181 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     6241 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     6301 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     6361 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
