LOCUS       sequence-226783-        8667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Iqsec1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8667)
  TITLE     Kahn expression plasmids
REFERENCE   2  (bases 1 to 8667)
  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"
COMMENT     Sequence Label: g13s pD47
FEATURES             Location/Qualifiers
     source          1..8667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        165..544
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        545..748
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     699..719
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     793..812
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        793..811
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    844..868
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3410..3434)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3445..3447,3448..3450,3451..4164)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3478..3506)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4101..4121
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4186..4203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4192..4416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4462..4890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4549..4568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4759..4780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4899..4919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4904..5233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5084..5219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5146..5165
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5300..6094
                     /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(5354..5373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5964..5983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6270..6391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6307..6326)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6361..6380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6440..6456)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6440..6456)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6453..6475)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6464..6480
                     /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(6488..6494,6495..6512,6513..6518))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6533..6554
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6671..6688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6842..7430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6922..6941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7601..8392,8393..8461))
                     /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     8224..8243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8462..8566)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8641..8660)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
       61 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      121 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      181 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      241 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      301 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      361 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      421 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      481 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      541 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      601 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      661 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      721 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      781 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt taagcttgat
      841 caaacaagtt tgtacaaaaa agcaggctta gaaggagata gaaccatgct agaacgaaag
      901 tatggggggc gcctggtaac ccgccatgcg gcccgcacca tccagacggc gtttcgccag
      961 taccagatga acaagaactt cgagcgcttg cgcagctcca tgtcagagaa ccgcatgtca
     1021 cgccggattg tgctgtccaa catgaggatg cagttctcct ttgaggggcc tgagaaagtg
     1081 cacagctcct acttcgaggg gaagcaggtc tcagtgacta acgacggctc ccagctggga
     1141 gccctggtgt cccctgagtg tggtgacctc agcgagccca ccaccctcaa gtctccggcc
     1201 ccctccagtg actttgcgga cgccatcacc gagctggagg acgccttctc taggcaagtg
     1261 aaatcactgg ccgagtccat cgacgatgcc ctcaactgcc gcagcctgca cactgaggag
     1321 gcaccggccc tggatgcggc gcgggcccgg gacaccgaac cccagacagc cctgcacggc
     1381 atggaccacc gcaaactgga cgagatgacg gcctcgtaca gtgatgtcac cctgtacatc
     1441 gatgaggagg agctgtcgcc ccctctgccc ctctcgcagg caggggaccg gccgtccagc
     1501 accgagtcgg acctgcggct acgggctggg ggcgcagccc cagactactg ggccctggcc
     1561 cacaaagagg acaaggctga cacggacacg agctgccgga gcacgccgtc gctggagcgg
     1621 caggagcagc ggctgcgggt ggagcatctg ccgctgctca ccatcgagcc acccagcgac
     1681 agctctgtgg accttagtga ccgctcggag cgggggtcac tcaagaggca gagtgcttac
     1741 gagcgcagcc ttggcgggca gcagggcagt cccaagcatg gtccccacag cggcgccccc
     1801 aagagcctcc cccgggagga gcctgagttg cggccccggc cccccaggcc cctggacagc
     1861 cacttggcca tcaatggctc agccaaccgg cagagcaagt ctgagtcgga ctactcagac
     1921 ggtgacaatg acagcatcaa cagcacgtcc aactccaacg ataccatcaa ctgcagctcc
     1981 gagtcatcgt cccgtgacag cctgcgggag cagacgctca gcaagcagac ctaccacaag
     2041 gaggcccgca acagctggga ctcgcctgcc tttagcaacg atgtcatccg caagaggcac
     2101 taccgcatcg gcctgaacct cttcaacaag aagcctgaga agggagtcca gtacctcatc
     2161 gagcgtggct ttgtgcccga cacgcccgtc ggggtggccc acttcctgct gcagcgcaag
     2221 ggcctcagcc ggcagatgat cggcgagttc ctgggcaacc ggcagaagca gttcaaccgt
     2281 gacgtgctcg actgcgtcgt ggacgagatg gacttctcta ccatggagct ggatgaggcc
     2341 ctcaggaaat tccaggcgca catccgtgtc caaggggagg ctcagaaagt ggagcggctc
     2401 atagaggcgt tcagccagcg ctactgcatc tgcaaccctg gggtggtgcg gcaattccgg
     2461 aacccagaca ccattttcat cctggccttc gccatcatcc tgctgaacac cgacatgtac
     2521 agccccaatg tcaagcccga gcggaaaatg aagctagagg acttcatcaa gaacctccga
     2581 ggtgtggacg atggtgagga cattccccgt gagatgctga tggggatcta tgaacggatc
     2641 cgtaagcgag agctaaagac caatgaggac catgtgtccc aggtgcagaa ggtggagaag
     2701 ctcattgtgg ggaaaaagcc gatcggatcc ctgcatcccg ggctcggctg tgtgctctct
     2761 ctgccccacc gtcggttggt ctgctactgc cggctctttg aggttccaga cccaaacaag
     2821 ccccagaaac tcggactaca ccagcgagaa atcttcctgt tcaacgacct cctggtggtc
     2881 accaagatct tccagaagaa gaagaactcg gtgacgtaca gcttccgaca gtccttctcc
     2941 ttgtacggca tgcaggtcct gctcttcgag aaccagtact accccaatgg catccggctc
     3001 acctcgtctg tccccggagc agatatcaaa gtgttaataa acttcaacgc ccccaaccct
     3061 caagaccgga agaaattcac cgatgacctg cgggagtcca ttgcggaagt ccaagagatg
     3121 gagaagcaca ggatagagtc ggagctcgag aagcagaaag gcgtcgtgcg gcccagcatg
     3181 tcccagtgct ctagcctcaa aaaggagtcg ggcaacggaa cactgagccg ggcctgcctg
     3241 gacgacagct atgccagcgg tgagggcctc aagcgcagcg ccctcagcag ctccctgcgg
     3301 gacctctcgg aagccgggaa gcgagggcgt cgcagcagtg cgggatcgct agagagcaat
     3361 gtggaatttc agcctttcga gccactgcag ccgtcagtgc tgtgctccga cccagctttc
     3421 ttgtacaaag tggttcgatc tagaatggct agcaaaggag aagaactttt cactggagtt
     3481 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
     3541 gagggtgaag gtgatgctac atacggaaag cttaccctta aatttatttg cactactgga
     3601 aaactacctg ttccatggcc aacacttgtc actactttct cttatggtgt tcaatgcttt
     3661 tcccgttatc cggatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt
     3721 tatgtacagg aacgcactat atctttcaaa gatgacggga actacaagac gcgtgctgaa
     3781 gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaaaggtat tgattttaaa
     3841 gaagatggaa acattctcgg acacaaactc gagtacaact ataactcaca caatgtatac
     3901 atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacatt
     3961 gaagatggat ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     4021 cctgtccttt taccagacaa ccattacctg tcgacacaat ctgccctttc gaaagatccc
     4081 aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat
     4141 ggcatggatg agctctacaa ataatgaatt aaacccgctg atcagcctcg actgtgcctt
     4201 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4261 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4321 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4381 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4441 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4501 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4561 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4621 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4681 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4741 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4801 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4861 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4921 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4981 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5041 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5101 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5161 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5221 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     5281 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5341 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5401 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5461 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5521 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5581 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5641 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5701 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5761 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5821 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5881 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5941 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6001 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6061 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     6121 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6181 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6241 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6301 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6361 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6421 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6481 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6541 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6601 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7261 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7501 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7561 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7681 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8461 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8521 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8581 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     8641 tacaatctgc tctgatgccg catagtt
//
