LOCUS       sequence-351672-        7438 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 7438)
  TITLE     eIF2-alpha constructs
REFERENCE   2  (bases 1 to 7438)
  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 1
FEATURES             Location/Qualifiers
     source          1..7438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2605..2623)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2606..2623)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2643..2660)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2649..2873
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2919..3347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3006..3025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3216..3237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3356..3376)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3361..3690
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3541..3676
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3603..3622
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5111..5232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5148..5167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5202..5221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5281..5297)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5281..5297)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5294..5316)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5305..5321
                     /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(5329..5335,5336..5353,5354..5359))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5374..5395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5512..5529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5683..6271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5763..5782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6442..7233,7234..7302))
                     /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     7065..7084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7303..7407)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttccggggaa gcaagtctgg
      961 tctctgtgat tgaagaagtc ggctctgggc tccagtgcgg gaatcacaca catacctcag
     1021 aatgccgggt ctaagttgta gattttatca acacaaattt cctgaggtgg aagatgtagt
     1081 gatggtgaat gtcagatcca ttgctgaaat gggggcttat gtcagcttgc tggaatacaa
     1141 caacattgaa ggcatgattc ttcttagtga attatccaga aggcgtatcc gttctatcaa
     1201 caaactcatc cgaattggca ggaatgagtg tgtggttgtc attagggtgg acaaagaaaa
     1261 aggatatatt gatttgtcaa aaagaagagt ttctccagag gaagcaatca aatgtgaaga
     1321 caaattcaca aaatccaaaa ctgtttatag cattcttcgt catgttgctg aggtgttaga
     1381 atacaccaag gatgagcagc tggaaagcct attccagagg actgcctggg tctttgatga
     1441 caagtacaag agacctggat atggtgccta tgatgcattt aagcatgcag tctcagaccc
     1501 atctattttg gatagtttag atttgaatga agatgaacgg gaagtactca ttaataatat
     1561 taataggcgc ttgaccccac aggctgtcaa aattcgagca gatattgaag tggcttgtta
     1621 tggttatgaa ggcattgatg ctgtaaaaga agccctaaga gcaggtttga attgttctac
     1681 agaaaacatg cccattaaga ttaatctaat agctcctcct cggtatgtaa tgactacgac
     1741 aaccctggag agaacagaag gcctttctgt cctcagtcaa gctatggctg ttatcaaaga
     1801 gaagattgag gaaaagaggg gtgtgttcaa tgttcaaatg gagcccaaag tggtcacaga
     1861 tacagatgag actgaacttg cgaggcagat ggagaggctt gaaagagaaa atgccgaagt
     1921 ggatggagat gatgatgcag aagaaatgga agccaaagct gaagattaac tttgtgggaa
     1981 acagagtcca atttaaggaa cacagagcag cgcttcctgg ctgtaaatcc tagacttgaa
     2041 agttttccag tattgaaaac ttcaaagctg aatatttttt atttctaagt atttaaatgt
     2101 tctaacagat cagaacatga aatgccctcc taaatgtcag ctgttgtcac acagtagctc
     2161 caacactttg agcattttta agggagtggc ctcatttcac tagagacaaa tctttaagaa
     2221 tagttctaaa attgggcttg tgatttccat ttctgatgtc tccagattgg cacccctttc
     2281 tagttcaatg cctcacgaga tttgccaggg gcatccaagg caaacaatcc caatctttct
     2341 atataaaatg tattcaagca aacatcaaat aaatttctgg gatatttaac tataggcttc
     2401 ttccttcttg tcaccagtta aaagcatttt aatactaaga ccctaattct tttatcttta
     2461 ttttagtctt gatgtggaac tgtaggagca ggtgaataaa ggatctctat aacagatcct
     2521 ttcaaaagaa gagttttaga gaaaataaat ttaactttaa ccacagtgaa agttgacctc
     2581 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     2641 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2701 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2761 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2821 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2881 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2941 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3001 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3061 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3121 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3181 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3241 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3301 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3361 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3421 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3481 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3541 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3601 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3661 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gctcgactag agatccctcg
     3721 acctcgagat ccattgtgct ctaaagccta agatgagctt tccatgtaaa tttgtagcca
     3781 gcttccttct gattttcaat gtttcttcca aaggtgcagt ctccaaagag attacgaatg
     3841 ccttggaaac ctggggtgcc ttgggtcagg acatcaactt ggacattcct agttttcaaa
     3901 tgagtgatga tattgacgat ataaaatggg aaaaaacttc agacaagaaa aagattgcac
     3961 aattcagaaa agagaaagag actttcaagg aaaaagatac atataagcta tttaaaaatg
     4021 gaactctgaa aattaagcat ctgaagaccg atgatcagga tatctacaag gtatcaatat
     4081 atgatacaaa aggaaaaaat gtgttggaaa aaatatttga tttgaagatt caagagaggg
     4141 tctcaaaacc aaagatctcc tggacttgta tcaacacaac cctgacctgt gaggtaatga
     4201 atggaactga ccccgaatta aacctgtatc aagatgggaa acatctaaaa ctttctcaga
     4261 gggtcatcac acacaagtgg accaccagcc tgagtgcaaa attcaagtgc acagcaggga
     4321 acaaagtcag caaggaatcc agtgtcgagc ctgtcagctg tccagagaaa ggtctggaca
     4381 tctatctcat cattggcata tgtggaggag gcagcctctt gatggtcttt gtggcactgc
     4441 tcgttttcta tatcaccaaa aggaaaaaac agaggagtcg gagaaatgat gaggagctgg
     4501 agacaagagc ccacagagta gctactgaag aaaggggccg gaagccccaa caaattccag
     4561 cttcaacccc tcagaatcca gcaacttccc aacatcctcc tccaccacct ggtcatcgtt
     4621 cccaggcacc tagtcatcgt cccccgcctc ctggacaccg tgttcagcac cagcctcaga
     4681 agaggcctcc tgctccgtcg ggcacacaag ttcaccagca gaaaggcccg cccctcccca
     4741 gacctcgagt tcagccaaaa cctccccatg gggcagcaga aaactcattg tccccttcct
     4801 ctaattaaaa aagatagaaa ctgtcttttt caataaaaag cactgtggat ttctgccctc
     4861 ctgatgtgca tatccgtact tccatgaggt gttttctgtg tgcagaacat tgtcacctcc
     4921 tgaggctgtg ggccacagcc acctctgcat cttcgaaatg accgaccaag cgacgcccaa
     4981 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5041 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5101 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5161 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5221 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5281 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5341 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5401 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5461 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5521 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5581 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5641 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5701 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5761 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5821 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5881 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5941 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6001 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6061 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6121 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6181 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6241 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6301 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6361 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6421 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6481 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6541 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6601 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6661 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6721 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6781 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6841 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6901 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6961 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7021 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7081 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7141 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7201 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7261 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7381 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
