LOCUS       sequence-358561-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  TITLE     eIF2-alpha constructs
REFERENCE   2  (bases 1 to 7420)
  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: eIF2a 3
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(74..91)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(245..833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(325..344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1004..1795,1796..1864))
                     /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     1627..1646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1865..1969)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2044..2063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2235..2614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2615..2818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2769..2789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2863..2882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2863..2881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4561..4577)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4561..4577)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4587..4605)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4588..4605)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4625..4642)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4631..4855
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4901..5329
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4988..5007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5198..5219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5338..5358)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5343..5672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5523..5658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5585..5604
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    7093..7214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7130..7149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7184..7203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7263..7279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7263..7279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7276..7298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7287..7303
                     /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(7311..7317,7318..7335,7336..7341))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7356..7377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
       61 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1081 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1141 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1201 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1261 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1321 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1381 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1441 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1501 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1561 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1621 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1681 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1741 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1801 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1861 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1921 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1981 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     2041 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     2101 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     2161 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     2221 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     2281 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2341 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2401 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2461 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     2521 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     2581 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     2641 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2701 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     2761 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     2821 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     2881 ggagacccaa gcttgatatc gaattccggg gaagcaagtc tggtctctgt gattgaagaa
     2941 gtcggctctg ggctccagtg cgggaatcac acacatacct cagaatgccg ggtctaagtt
     3001 gtagatttta tcaacacaaa tttcctgagg tggaagatgt agtgatggtg aatgtcagat
     3061 ccattgctga aatgggggct tatgtcagct tgctggaata caacaacatt gaaggcatga
     3121 ttcttcttag tgaactcgac agaaggcgta tccgttctat caacaaactc atccgaattg
     3181 gcaggaatga gtgtgtggtt gtcattaggg tggacaaaga aaaaggatat attgatttgt
     3241 caaaaagaag agtttctcca gaggaagcaa tcaaatgtga agacaaattc acaaaatcca
     3301 aaactgttta tagcattctt cgtcatgttg ctgaggtgtt agaatacacc aaggatgagc
     3361 agctggaaag cctattccag aggactgcct gggtctttga tgacaagtac aagagacctg
     3421 gatatggtgc ctatgatgca tttaagcatg cagtctcaga cccatctatt ttggatagtt
     3481 tagatttgaa tgaagatgaa cgggaagtac tcattaataa tattaatagg cgcttgaccc
     3541 cacaggctgt caaaattcga gcagatattg aagtggcttg ttatggttat gaaggcattg
     3601 atgctgtaaa agaagcccta agagcaggtt tgaattgttc tacagaaaac atgcccatta
     3661 agattaatct aatagctcct cctcggtatg taatgactac gacaaccctg gagagaacag
     3721 aaggcctttc tgtcctcagt caagctatgg ctgttatcaa agagaagatt gaggaaaaga
     3781 ggggtgtgtt caatgttcaa atggagccca aagtggtcac agatacagat gagactgaac
     3841 ttgcgaggca gatggagagg cttgaaagag aaaatgccga agtggatgga gatgatgatg
     3901 cagaagaaat ggaagccaaa gctgaagatt aactttgtgg gaaacagagt ccaatttaag
     3961 gaacacagag cagcgcttcc tggctgtaaa tcctagactt gaaagttttc cagtattgaa
     4021 aacttcaaag ctgaatattt tttatttcta agtatttaaa tgttctaaca gatcagaaca
     4081 tgaaatgccc tcctaaatgt cagctgttgt cacacagtag ctccaacact ttgagcattt
     4141 ttaagggagt ggcctcattt cactagagac aaatctttaa gaatagttct aaaattgggc
     4201 ttgtgatttc catttctgat gtctccagat tggcacccct ttctagttca atgcctcacg
     4261 agatttgcca ggggcatcca aggcaaacaa tcccaatctt tctatataaa atgtattcaa
     4321 gcaaacatca aataaatttc tgggatattt aactataggc ttcttccttc ttgtcaccag
     4381 ttaaaagcat tttaatacta agaccctaat tcttttatct ttattttagt cttgatgtgg
     4441 aactgtagga gcaggtgaat aaaggatctc tataacagat cctttcaaaa gaagagtttt
     4501 agagaaaata aatttaactt taaccacagt gaaagttgac ctcgagcatg cagcccgggg
     4561 gatccactag ttctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     4621 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4681 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4741 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4801 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4861 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4921 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4981 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5041 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5101 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5161 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5221 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5281 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5341 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5401 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5461 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5521 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5581 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5641 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggctcgact agagatccct
     5701 cgacctcgag atccattgtg ctctaaagcc taagatgagc tttccatgta aatttgtagc
     5761 cagcttcctt ctgattttca atgtttcttc caaaggtgca gtctccaaag agattacgaa
     5821 tgccttggaa acctggggtg ccttgggtca ggacatcaac ttggacattc ctagttttca
     5881 aatgagtgat gatattgacg atataaaatg ggaaaaaact tcagacaaga aaaagattgc
     5941 acaattcaga aaagagaaag agactttcaa ggaaaaagat acatataagc tatttaaaaa
     6001 tggaactctg aaaattaagc atctgaagac cgatgatcag gatatctaca aggtatcaat
     6061 atatgataca aaaggaaaaa atgtgttgga aaaaatattt gatttgaaga ttcaagagag
     6121 ggtctcaaaa ccaaagatct cctggacttg tatcaacaca accctgacct gtgaggtaat
     6181 gaatggaact gaccccgaat taaacctgta tcaagatggg aaacatctaa aactttctca
     6241 gagggtcatc acacacaagt ggaccaccag cctgagtgca aaattcaagt gcacagcagg
     6301 gaacaaagtc agcaaggaat ccagtgtcga gcctgtcagc tgtccagaga aaggtctgga
     6361 catctatctc atcattggca tatgtggagg aggcagcctc ttgatggtct ttgtggcact
     6421 gctcgttttc tatatcacca aaaggaaaaa acagaggagt cggagaaatg atgaggagct
     6481 ggagacaaga gcccacagag tagctactga agaaaggggc cggaagcccc aacaaattcc
     6541 agcttcaacc cctcagaatc cagcaacttc ccaacatcct cctccaccac ctggtcatcg
     6601 ttcccaggca cctagtcatc gtcccccgcc tcctggacac cgtgttcagc accagcctca
     6661 gaagaggcct cctgctccgt cgggcacaca agttcaccag cagaaaggcc cgcccctccc
     6721 cagacctcga gttcagccaa aacctcccca tggggcagca gaaaactcat tgtccccttc
     6781 ctctaattaa aaaagataga aactgtcttt ttcaataaaa agcactgtgg atttctgccc
     6841 tcctgatgtg catatccgta cttccatgag gtgttttctg tgtgcagaac attgtcacct
     6901 cctgaggctg tgggccacag ccacctctgc atcttcgaaa tgaccgacca agcgacgccc
     6961 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     7021 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     7081 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7141 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7201 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     7261 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7321 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7381 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
//
