LOCUS       sequence-485104-        6613 bp    DNA     circular SYN 02-SEP-2025
DEFINITION  Expresses full length NRL, driven by EF1-alpha promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6613)
  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 6613)
  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-FL-NRL
FEATURES             Location/Qualifiers
     source          1..6613
                     /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(2621..2638)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2627..2851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2897..3325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2984..3003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3194..3215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3334..3354)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3339..3668
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3519..3654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3581..3600
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3716..3763
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3782..4156
                     /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    4286..4407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4323..4342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4377..4396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4456..4472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4456..4472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4469..4491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4480..4496
                     /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(4504..4510,4511..4528,4529..4534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4549..4570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4687..4704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4858..5446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4938..4957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5617..6408,6409..6477))
                     /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     6240..6259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6478..6582)
                     /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 tccgaaacca tggccctgcc ccccagcccc
     1861 ctggccatgg aatatgtcaa tgactttgac ttgatgaagt ttgaggtaaa gcgggaaccc
     1921 tctgagggcc gacctggccc ccctacagcc tcactgggct ccacacctta cagctcagtg
     1981 cctccttcac ccaccttcag tgaaccaggc atggtggggg caaccgaggg cacccggcca
     2041 ggcctggagg agctgtactg gctggctacc ctgcagcagc agctgggggc tggggaggca
     2101 ttggggctga gtcctgaaga ggccatggag ctgctgcagg gtcagggccc agtccctgtt
     2161 gatgggcccc atggctacta cccagggagc ccagaggaga caggagccca gcacgtccag
     2221 ctggcagagc ggttttccga cgcggcgctg gtctcgatgt ctgtgcggga gctaaaccgg
     2281 cagctgcggg gctgcgggcg cgacgaggcg ctgcggctga agcagaggcg ccgcacgctg
     2341 aagaaccgcg gctacgcgca ggcctgtcgc tccaagcggc tgcagcagcg gcgcgggctg
     2401 gaggccgagc gcgcccgcct ggccgcccag ctggacgcgc tgcgggccga ggtggcccgc
     2461 ctggcccggg agcgcgatct ctacaaggct cgctgtgacc ggctaacctc gagcggcccc
     2521 gggtccgggg acccctccca cctcttcctc tgagccgttc agagcacctt gtggtggcgg
     2581 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     2641 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2701 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2761 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2821 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2881 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3241 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3301 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3361 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3421 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3481 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3541 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3601 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3661 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     3721 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3781 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     3841 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     3901 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     3961 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     4021 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     4081 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     4141 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     4201 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4261 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4321 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4381 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4441 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4501 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4561 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4621 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5041 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5281 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6601 gccacctgac gtc
//
