LOCUS       Exported                6185 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses SLC39A14/ZIP14 wild-type in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    SLC39A14-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6185)
  AUTHORS   Liu C, Hutchens S, Jursa T, Shawlot W, Polishchuk EV, Polishchuk RS,
            Dray BK, Gore AC, Aschner M, Smith DR, Mukhopadhyay S
  TITLE     Hypothyroidism induced by loss of the manganese efflux transporter 
            SLC30A10 may be explained by reduced thyroxine production.
  JOURNAL   J Biol Chem. 2017 Oct 6;292(40):16605-16615. doi: 
            10.1074/jbc.M117.804989. Epub 2017 Aug 31.
  PUBMED    28860195
REFERENCE   2  (bases 1 to 6185)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6185
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2065..2074
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2071..2790
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2071..2787
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2116..2137)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2377..2396)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2724..2745
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2913..3034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2950..2969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3004..3023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3041..3496)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3178..3199)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3390..3409
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3523..3627
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3629..3986
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3652..3672)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3837..3972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3899..3918
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4021..4815
                     /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(4075..4094)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4685..4704
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5003..5022)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5047..5094
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5423..6011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5912..5931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacga taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttgtc gacggtacca tgaagctgct gctgctgcac
      601 ccggccttcc agagctgcct cctgctgacc ctgcttggct tatggagaac cacccctgag
      661 gctcacgctt catccctggg tgcaccagct atcagcgctg cctccttcct gcaggatcta
      721 atacatcggt atggcgaggg tgacagcctc actctgcagc agctgaaggc cctactcaac
      781 cacctggatg tgggagtggg ccggggtaat gtcacccagc acgtgcaagg acacaggaac
      841 ctctccacgt gctttagttc tggagacctc ttcactgccc acaatttcag cgagcagtcg
      901 cggattggga gcagcgagct ccaggagttc tgccccacca tcctccagca gctggattcc
      961 cgggcctgca cctcggagaa ccaggaaaac gaggagaatg agcagacgga ggaggggcgg
     1021 ccaagcgctg ttgaagtgtg gggatacggt ctcctctgtg tgaccgtcat ctccctctgc
     1081 tccctcctgg gggccagcgt ggtgcccttc atgaagaaga ccttttacaa gaggctgctg
     1141 ctctacttca tagctctggc gattggaacc ctctactcca acgccctctt ccagctcatc
     1201 ccggaggcat ttggtttcaa ccctctggaa gattattatg tctccaagtc tgcagtggtg
     1261 tttgggggct tttatctttt ctttttcaca gagaagatct tgaagattct tcttaagcag
     1321 aaaaatgagc atcatcatgg acacagccat tatgcctctg agtcgcttcc ctccaagaag
     1381 gaccaggagg agggggtgat ggagaagctg cagaacgggg acctggacca catgattcct
     1441 cagcactgca gcagtgagct ggacggcaag gcgcccatgg tggacgagaa ggtcattgtg
     1501 ggctcgctct ctgtgcagga cctgcaggct tcccagagtg cttgctactg gctgaaaggt
     1561 gtccgctact ctgatatcgg cactctggcc tggatgatca ctctgagcga cggcctccat
     1621 aatttcatcg atggcctggc catcggtgct tccttcactg tgtcagtttt ccaaggcatc
     1681 agcacctcgg tggccatcct ctgtgaggag ttcccacatg agctaggaga ctttgtcatc
     1741 ctgctcaacg ctgggatgag catccaacaa gctctcttct tcaacttcct ttctgcctgc
     1801 tgctgctacc tgggtctggc ctttggcatc ctggccggca gccacttctc tgccaactgg
     1861 atttttgcgc tagctggagg aatgttcttg tatatttctc tggctgatat gttccctgag
     1921 atgaatgagg tctgtcaaga ggatgaaagg aagggcagca tcttgattcc atttatcatc
     1981 cagaacctgg gcctcctgac tggattcacc atcatggtgg tcctcaccat gtattcagga
     2041 cagatccaga ttgggggatc catcgccacc atggtgagca agggcgagga gctgttcacc
     2101 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2161 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     2221 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2281 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2341 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2401 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2461 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2521 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2581 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2641 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2701 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2761 actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc
     2821 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2881 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2941 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3001 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     3061 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca atatgccgaa
     3121 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3181 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3241 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3301 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3361 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3421 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3481 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3541 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3601 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3661 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3721 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3781 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3841 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3901 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3961 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4021 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4081 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4141 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4201 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4261 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4321 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4381 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4441 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4501 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4561 ggcgaagatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4621 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4681 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4741 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4801 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4861 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4921 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4981 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5041 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5101 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggtc
     5161 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5221 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5281 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5341 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5401 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5461 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5521 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5581 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5641 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5701 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5761 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5821 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5881 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5941 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6001 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6061 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6121 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     6181 ttccg
//
