LOCUS       sequence-229779-        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 2
FEATURES             Location/Qualifiers
     source          1..7438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     861..880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        861..879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2603..2621)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2604..2621)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2641..2658)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2647..2871
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2917..3345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3004..3023)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3214..3235
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3354..3374)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3359..3688
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3539..3674
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3601..3620
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5109..5230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5146..5165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5200..5219
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5279..5295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5279..5295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5292..5314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5303..5319
                     /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(5327..5333,5334..5351,5352..5357))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5372..5393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5510..5527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5681..6269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5761..5780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6440..7231,7232..7300))
                     /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     7063..7082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7301..7405)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga
      901 gctcggatcc actagtaacg gccgccagtg tgctggaatt ccggggaagc aagtctggtc
      961 tctgtgattg aagaagtcgg ctctgggctc cagtgcggga atcacacaca tacctcagaa
     1021 tgccgggtct aagttgtaga ttttatcaac acaaatttcc tgaggtggaa gatgtagtga
     1081 tggtgaatgt cagatccatt gctgaaatgg gggcttatgt cagcttgctg gaatacaaca
     1141 acattgaagg catgattctt cttagtgaat tggccagaag gcgtatccgt tctatcaaca
     1201 aactcatccg aattggcagg aatgagtgtg tggttgtcat tagggtggac aaagaaaaag
     1261 gatatattga tttgtcaaaa agaagagttt ctccagagga agcaatcaaa tgtgaagaca
     1321 aattcacaaa atccaaaact gtttatagca ttcttcgtca tgttgctgag gtgttagaat
     1381 acaccaagga tgagcagctg gaaagcctat tccagaggac tgcctgggtc tttgatgaca
     1441 agtacaagag acctggatat ggtgcctatg atgcatttaa gcatgcagtc tcagacccat
     1501 ctattttgga tagtttagat ttgaatgaag atgaacggga agtactcatt aataatatta
     1561 ataggcgctt gaccccacag gctgtcaaaa ttcgagcaga tattgaagtg gcttgttatg
     1621 gttatgaagg cattgatgct gtaaaagaag ccctaagagc aggtttgaat tgttctacag
     1681 aaaacatgcc cattaagatt aatctaatag ctcctcctcg gtatgtaatg actacgacaa
     1741 ccctggagag aacagaaggc ctttctgtcc tcagtcaagc tatggctgtt atcaaagaga
     1801 agattgagga aaagaggggt gtgttcaatg ttcaaatgga gcccaaagtg gtcacagata
     1861 cagatgagac tgaacttgcg aggcagatgg agaggcttga aagagaaaat gccgaagtgg
     1921 atggagatga tgatgcagaa gaaatggaag ccaaagctga agattaactt tgtgggaaac
     1981 agagtccaat ttaaggaaca cagagcagcg cttcctggct gtaaatccta gacttgaaag
     2041 ttttccagta ttgaaaactt caaagctgaa tattttttat ttctaagtat ttaaatgttc
     2101 taacagatca gaacatgaaa tgccctccta aatgtcagct gttgtcacac agtagctcca
     2161 acactttgag catttttaag ggagtggcct catttcacta gagacaaatc tttaagaata
     2221 gttctaaaat tgggcttgtg atttccattt ctgatgtctc cagattggca cccctttcta
     2281 gttcaatgcc tcacgagatt tgccaggggc atccaaggca aacaatccca atctttctat
     2341 ataaaatgta ttcaagcaaa catcaaataa atttctggga tatttaacta taggcttctt
     2401 ccttcttgtc accagttaaa agcattttaa tactaagacc ctaattcttt tatctttatt
     2461 ttagtcttga tgtggaactg taggagcagg tgaataaagg atctctataa cagatccttt
     2521 caaaagaaga gttttagaga aaataaattt aactttaacc acagtgaaag ttgacctcga
     2581 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
     2641 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2701 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2761 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2821 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2881 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2941 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3001 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3061 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3121 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3181 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3241 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3301 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3361 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3421 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3481 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3541 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3601 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     3661 cttttttgga ggcctaggct tttgcaaaaa gctcccgggc tcgactagag atccctcgac
     3721 ctcgagatcc attgtgctct aaagcctaag atgagctttc catgtaaatt tgtagccagc
     3781 ttccttctga ttttcaatgt ttcttccaaa ggtgcagtct ccaaagagat tacgaatgcc
     3841 ttggaaacct ggggtgcctt gggtcaggac atcaacttgg acattcctag ttttcaaatg
     3901 agtgatgata ttgacgatat aaaatgggaa aaaacttcag acaagaaaaa gattgcacaa
     3961 ttcagaaaag agaaagagac tttcaaggaa aaagatacat ataagctatt taaaaatgga
     4021 actctgaaaa ttaagcatct gaagaccgat gatcaggata tctacaaggt atcaatatat
     4081 gatacaaaag gaaaaaatgt gttggaaaaa atatttgatt tgaagattca agagagggtc
     4141 tcaaaaccaa agatctcctg gacttgtatc aacacaaccc tgacctgtga ggtaatgaat
     4201 ggaactgacc ccgaattaaa cctgtatcaa gatgggaaac atctaaaact ttctcagagg
     4261 gtcatcacac acaagtggac caccagcctg agtgcaaaat tcaagtgcac agcagggaac
     4321 aaagtcagca aggaatccag tgtcgagcct gtcagctgtc cagagaaagg tctggacatc
     4381 tatctcatca ttggcatatg tggaggaggc agcctcttga tggtctttgt ggcactgctc
     4441 gttttctata tcaccaaaag gaaaaaacag aggagtcgga gaaatgatga ggagctggag
     4501 acaagagccc acagagtagc tactgaagaa aggggccgga agccccaaca aattccagct
     4561 tcaacccctc agaatccagc aacttcccaa catcctcctc caccacctgg tcatcgttcc
     4621 caggcaccta gtcatcgtcc cccgcctcct ggacaccgtg ttcagcacca gcctcagaag
     4681 aggcctcctg ctccgtcggg cacacaagtt caccagcaga aaggcccgcc cctccccaga
     4741 cctcgagttc agccaaaacc tccccatggg gcagcagaaa actcattgtc cccttcctct
     4801 aattaaaaaa gatagaaact gtctttttca ataaaaagca ctgtggattt ctgccctcct
     4861 gatgtgcata tccgtacttc catgaggtgt tttctgtgtg cagaacattg tcacctcctg
     4921 aggctgtggg ccacagccac ctctgcatct tcgaaatgac cgaccaagcg acgcccaacc
     4981 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5041 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5101 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5161 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5221 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5281 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5341 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6121 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcga
//
