LOCUS       sequence-485105-        5899 bp    DNA     circular SYN 02-SEP-2025
DEFINITION  Empty pcDNA4-C backbone with EF1-alpha promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5899)
  AUTHORS   Shayler DWH, Stachelek K, Cambier L, Lee S, Bai J, Reid MW, 
            Weisenberger DJ, Bhat B, Aparicio JG, Kim Y, Singh M, Bay M, 
            Thornton ME, Doyle EK, Fouladian Z, Erberich SG, Grubbs BH, 
            Bonaguidi MA, Craft CM, Singh HP, Cobrinik D
  TITLE     Identification and characterization of early human photoreceptor 
            states and cell-state-specific retinoblastoma-related features
  JOURNAL   eLife 13:RP101918
REFERENCE   2  (bases 1 to 5899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife 
            13:RP101918"
COMMENT     Sequence Label: pcDNA4-C-EF1-alpha
FEATURES             Location/Qualifiers
     source          1..5899
                     /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"
     promoter        378..1556
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          609..1547
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1504..1524
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1623..1642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1623..1641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1677..1839
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     primer_bind     complement(1907..1924)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1913..2137
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2183..2611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2270..2289)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2480..2501
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2620..2640)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2625..2954
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2805..2940
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2867..2886
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3002..3049
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3068..3442
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3572..3693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3609..3628)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3663..3682
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3742..3758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3742..3758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3755..3777)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3766..3782
                     /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(3790..3796,3797..3814,3815..3820))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3835..3856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3973..3990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4144..4732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4224..4243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4903..5694,5695..5763))
                     /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     5526..5545
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5764..5868)
                     /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 tagcaaattg ggacattgat tattgactag gcttttgcaa aaagctttgc
      301 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      361 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      421 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      481 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      541 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      601 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      661 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
      721 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
      781 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
      841 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
      901 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
      961 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1021 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1081 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1141 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1201 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1261 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1321 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1381 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1441 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1501 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa
     1561 ttagcttggt accgtcctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     1621 attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt aagcttagcg
     1681 cagaggcttg gggcagccga gcggcagcca ggccccggcc cgggcctcgg ttccagaagg
     1741 gagaggagcc cgccaaggcg cgcaagagag cgggctgcct cgcagtccga gccggagagg
     1801 gagcgcgagc cgcgccggcc ccggacggcc tccgaaaccg ccgttcagag caccttgtgg
     1861 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1921 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1981 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2041 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2101 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2161 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2221 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2281 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2341 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2401 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2461 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2521 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2581 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2641 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2701 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2761 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2821 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2881 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2941 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     3001 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     3061 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     3121 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     3181 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     3241 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     3301 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     3361 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     3421 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     3481 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3541 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3601 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3661 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3721 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3781 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3841 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3901 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3961 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4021 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4081 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4321 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4561 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4681 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4801 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4861 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4921 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4981 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5041 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5101 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5161 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5221 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5281 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5341 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5401 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5461 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5521 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5581 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5641 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5701 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5761 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5821 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5881 aaaagtgcca cctgacgtc
//
