LOCUS       sequence-473344-        5356 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express HiBiT-SLC35A2 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5356)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HiBiT-SLC35A2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2435..2453)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2490..2611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2501..2520)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2555..2574
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2856..3213
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2856..3047
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2879..2899)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3064..3199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3228..3275
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3307..4101
                     /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(3361..3380)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3971..3990
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4165..4213
                     /note="synthetic polyadenylation signal"
     rep_origin      4370..4915
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4816..4835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5032..5049
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatggctg
     1141 ccgtgggcgc aggaggaagt acggcagcac ctggtccggg cgcggtgtct gcaggcgctc
     1201 ttgaacctgg gactgcttca gccgcccacc gaagattgaa gtatatttct cttgctgttc
     1261 tcgtagtgca aaatgcttcc ctcatccttt ctataagata cgctcggacg ttgccgggcg
     1321 atcgattttt cgcaacaacc gcagtcgtca tggcagaggt ccttaaggga ttgacgtgcc
     1381 tgctccttct ctttgcgcaa aaacgaggaa atgtaaaaca cctggttctg tttctgcacg
     1441 aggcagtgct tgtccaatat gttgacacac tgaagttggc ggtacccagc ctcatttata
     1501 ctttgcaaaa taatcttcaa tatgtcgcta tatcaaattt gccggcagct acgttccaag
     1561 tcacctacca actgaaaatc ctgacaacgg cattgttctc cgttcttatg ctgaaccggt
     1621 ccttgtcccg actccaatgg gccagtctcc ttctcctctt tacaggggtc gccatagttc
     1681 aagctcaaca agcgggagga ggaggtcccc gccctctcga tcagaatccg ggagccgggc
     1741 tcgcggctgt ggtggcctcc tgccttagct ctgggttcgc gggtgtatat tttgagaaga
     1801 tcctgaaggg ttcttctggt agtgtctggt tgagaaatct gcagcttggc ctctttggga
     1861 cggccctcgg gttggtgggt ttgtggtggg cggagggcac cgcagtcgca actcgcggat
     1921 tcttttttgg atacacacca gctgtttggg gtgtcgtatt gaaccaagca tttggcggat
     1981 tgctcgtggc ggttgttgtg aaatacgcgg acaacatact gaagggattc gcgacctcct
     2041 tgtccatcgt attgtccacc gtcgcctcaa taagactctt cgggtttcat gtagatccgc
     2101 tctttgcctt gggtgctggg cttgtcatag gagcggtcta tttgtactcc ctgcctcggg
     2161 gagcagcgaa agccatagcc tctgcttccg cgtctgcctc cggtccgtgc gtccatcagc
     2221 aacctccggg acagccgccg cctccacagt tgtcctccca ccgcggtgac ctcataaccg
     2281 agcccttcct gccgaaattg ctgaccaaag tgaagggatc cgtttaaacg aattcgggct
     2341 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagctgatcc ggctgctaac
     2401 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2461 cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2521 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2581 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2641 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     2701 taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     2761 tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa
     2821 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     2881 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2941 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3001 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3061 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3121 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3181 ggaggctttt ttggaggcct aggcttttgc aaaaagctta attaactgtt gacaattaat
     3241 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg
     3301 cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3361 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3421 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3481 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3541 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3601 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3661 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3721 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3781 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3841 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3901 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3961 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4021 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4081 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4141 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     4201 ggttttttgt gtgaatcgat agcgataagg atccgctcat gcgatgataa gctgtcaaac
     4261 atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt gaagagataa
     4321 attgcactga aatctagaaa tattttatct gattaataag atgatcttct tgagatcgtt
     4381 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
     4441 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt
     4501 caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct
     4561 ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac
     4621 tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata
     4681 cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa
     4741 acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac
     4801 gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacca
     4861 ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg
     4921 ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc caggaaatct
     4981 ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca
     5041 gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac cggtgcagcc
     5101 ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca acatagtaag
     5161 ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac gcaccacccc
     5221 gtcatgagat tacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta
     5281 ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg
     5341 tagaggatcg agatct
//
